Methods of treating a disease or disorder associated with bruton&#39;s tyrosine kinase

ABSTRACT

The present invention provides methods of treating, stabilizing or lessening the severity or progression of a disease or disorder associated with BTK.

CROSS REFERENCE TO RELATED APPLICATIONS

The present application claims priority to U.S. provisional applicationNo. 61/870,720, filed Aug. 27, 2013, the entirety of which is herebyincorporated by reference.

FIELD OF THE INVENTION

The present invention provides methods of treating, stabilizing orlessening the severity or progression of a disease or disorderassociated with Bruton's Tyrosine Kinase (“BTK”).

BACKGROUND OF THE INVENTION

The search for new therapeutic agents has been greatly aided in recentyears by a better understanding of the structure of enzymes and otherbiomolecules associated with diseases. One important class of enzymesthat has been the subject of extensive study is protein kinases.

Protein kinases constitute a large family of structurally relatedenzymes that are responsible for the control of a variety of signaltransduction processes within the cell. Protein kinases are thought tohave evolved from a common ancestral gene due to the conservation oftheir structure and catalytic function. Almost all kinases contain asimilar 250-300 amino acid catalytic domain. The kinases may becategorized into families by the substrates they phosphorylate (e.g.,protein-tyrosine, protein-serine/threonine, lipids, etc.).

In general, protein kinases mediate intracellular signaling by effectinga phosphoryl transfer from a nucleoside triphosphate to a proteinacceptor that is involved in a signaling pathway. These phosphorylationevents act as molecular on/off switches that can modulate or regulatethe target protein biological function. These phosphorylation events areultimately triggered in response to a variety of extracellular and otherstimuli. Examples of such stimuli include environmental and chemicalstress signals (e.g., osmotic shock, heat shock, ultraviolet radiation,bacterial endotoxin, and H₂O₂), cytokines (e.g., interleukin-1 (IL-1)and tumor necrosis factor α (TNF-α)), and growth factors (e.g.,granulocyte macrophage-colony-stimulating factor (GM-CSF), andfibroblast growth factor (FGF)). An extracellular stimulus may affectone or more cellular responses related to cell growth, migration,differentiation, secretion of hormones, activation of transcriptionfactors, glucose metabolism, control of protein synthesis, andregulation of the cell cycle.

Many diseases are associated with abnormal cellular responses triggeredby protein kinase-mediated events as described above. These diseasesinclude, but are not limited to, autoimmune diseases, inflammatorydiseases, bone diseases, metabolic diseases, neurological andneurodegenerative diseases, cancer, cardiovascular diseases, allergiesand asthma, Alzheimer's disease, and hormone-related diseases.Accordingly, there remains a need to find protein kinase inhibitorsuseful as therapeutic agents.

SUMMARY OF THE INVENTION

Chronic lymphocytic leukemia (CLL) is a lymphoproliferative malignancycharacterized by progressive accumulation of morphologically mature butfunctionally incompetent lymphocytes in the blood, bone marrow, andlymphoid tissues. It affects mainly elderly individuals with the medianage at presentation of 65 to 70 years. Small lymphocytic lymphoma (SLL)and CLL are generally considered a different manifestation of the samedisease. While CLL is found in the blood and bone marrow, SLL presentsprimarily in the lymph nodes. The clinical course of CLL/SLL ranges fromindolent disease with long-term survival over 12 years to aggressivedisease with median survival of 2 years. The average age of diagnosiswith CLL/SLL is approximately 60 years.

Despite newly approved therapeutic agents and combination therapies,CLL/SLL remains an incurable disease and most patients eventuallyrelapse and/or die. Improved and novel combination treatments forsubjects with CLL/SLL requiring treatment remain an unmet medical need.

Bruton's tyrosine kinase (Btk) is a non-receptor tyrosine kinase withrestricted cellular expression largely limited to B-lymphocytes,monocytes, and mast cells or basophils. Btk is a critical component ofthe B-cell receptor (BCR) signaling network and is crucial for B-celldevelopment. Investigation has revealed that some B-cell malignancies,including diseases such as CLL/SLL, depend on BCR signaling, suggestingthat interruption of such signaling could be a promising therapeuticopportunity. Recently, clinical anti-tumor responses in various B-cellnon-Hodgkin's Lymphoma (NHL) and CLL/SLL have been reported with agentsthat inhibit spleen tyrosine kinase (Syk) and Btk, both components ofthe BCR signaling pathway.

United States published patent application number US 2010/0029610,published Feb. 4, 2010 (“the '610 publication,” the entirety of which ishereby incorporated herein by reference), describes certain2,4-disubstituted pyrimidine compounds which covalently and irreversiblyinhibit activity of one or more protein kinases, including BTK, a memberof TEC-kinases. Such compounds includeN-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamide,hereinafter referred to as Compound 1, which is designated as compoundnumber I-182 in the '610 publication. The synthesis of Compound 1 isdescribed in detail at Example 20 of the '610 publication Compound 1 isactive in a variety of assays and therapeutic models demonstratingcovalent, irreversible inhibition of BTK (in enzymatic and cellularassays). Notably, Compound 1 is a potent, selective, orally available,small molecule which was found to inhibit B-cell proliferation andactivation. Compound 1 is therefore useful for treating one or moredisorders associated with activity of BTK.

Accordingly, among other things, the present invention provides methodsof treating, stabilizing or lessening the severity or progression of oneor more diseases and conditions associated with BTK. In some aspects,the present invention provides methods of treating, stabilizing orlessening the severity or progression of one or more diseases andconditions associated with BTK comprising administering to a patient inneed thereof a pharmaceutically acceptable composition comprisingN-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamide(1):

In some embodiments, the present invention provides a method oftreating, stabilizing or lessening the severity or progression of one ormore diseases and conditions associated with BTK comprisingadministering to a patient in need thereof Compound 1 in combinationwith rituximab.

In some embodiments, the present invention provides a method oftreating, stabilizing or lessening the severity or progression of one ormore diseases and conditions associated with BTK comprisingadministering to a patient in need thereof a composition comprisingCompound 1 in combination with a composition comprising rituximab.

In some embodiments, provided methods comprise administering to apatient in need thereof Compound 1 in combination with rituximab,wherein Compound 1 is administered once a day. In some embodiments,provided methods comprise administering to a patient in need thereofCompound 1 in combination with rituximab, wherein Compound 1 isadministered twice a day. In some such embodiments, rituximab isadministered once during a 28-day cycle. Accordingly, in someembodiments, provided methods comprise administering to a patient inneed thereof Compound 1 in combination with rituximab, wherein Compound1 is administered twice a day and rituximab is administered once duringa 28-day cycle.

In some embodiments, the provided methods comprise administering to apatient in need thereof a composition comprising Compound 1 andrituximab.

In some embodiments, the provided methods comprise administering to apatient in need thereof Compound 1, rituximab, fludarabine andcyclophosphamide.

In some embodiments, the provided methods comprise administering to apatient in need thereof Compound 1, rituximab and bendamustine.

In some embodiments, the disease or condition associated with BTK isselected from chronic lymphocytic leukemia and small lymphocyticlymphoma.

In some embodiments, the present invention provides a method oftreating, stabilizing or lessening the severity or progression ofchronic lymphocytic leukemia (CLL), the method comprising administeringto a patient in need thereof Compound 1 in combination with rituximab.

In some embodiments, the present invention provides a method oftreating, stabilizing or lessening the severity or progression of smalllymphocytic lymphoma (SLL), the method comprising administering to apatient in need thereof Compound 1 in combination with rituximab.

In some embodiments, provided therapies comprise orally administering toa patient Compound 1 in combination with rituximab. In some embodiments,each of Compound 1 and rituximab is administered in the form of apharmaceutical formulation. In some embodiments, the pharmaceuticalformulation comprising Compound 1 is a capsule formulation. In someembodiments, the pharmaceutical formulation comprising rituximab is anintravenous (IV) formulation.

In some embodiments, the present invention also provides dosing regimensand protocols for administering to patients in need thereof Compound 1in combination with rituximab. Such methods, dosing regimens andprotocols for the administration of said combination are described infurther detail, below.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 presents the Response Assessments of patients enrolled in cohorts1 and 2 as of Oct. 16, 2013.

DETAILED DESCRIPTION OF THE INVENTION Definitions

As used herein, the term “antibody”, or grammatical variations thereof(i.e., antibodies), refers to polypeptide(s) capable of binding to anepitope. In some embodiments, an antibody is a full-length antibody. Insome embodiments, an antibody is less than full length (i.e., anantibody fragment) but includes at least one binding site. In some suchembodiments, the binding site comprises at least one, and preferably atleast two sequences with structure of antibody variable regions. In someembodiments, the term “antibody” encompasses any protein having abinding domain which is homologous or largely homologous to animmunoglobulin-binding domain. In particular embodiments, the term“antibody” encompasses polypeptides having a binding domain that showsat least 99% identity with an immunoglobulin-binding domain. In someembodiments, the antibody is any protein having a binding domain thatshows at least 70%, at least 80%, at least 85%, at least 90% or at least95% identity with an immunoglobulin-binding domain. Antibodypolypeptides in accordance with the present invention may be prepared byany available means, including, for example, isolation from a naturalsource or antibody library, recombinant production in or with a hostsystem, chemical synthesis, etc., or combinations thereof. In someembodiments, an antibody is monoclonal or polyclonal. In someembodiments, an antibody may be a member of any immunoglobulin class,including any of the human classes IgG, IgM, IgA, IgD and IgE. Incertain embodiments, an antibody is a member of the IgG immunoglobulinclass. In some embodiments, the term “antibody” refers to any derivativeof an antibody that possesses the ability to bind to an epitope ofinterest. In some embodiments, an antibody fragment comprises multiplechains that are linked together, for example, by disulfide linkages. Insome embodiments, an antibody is a human antibody. In some embodiments,an antibody is a humanized antibody. In some embodiments, humanizedantibodies include chimeric immunoglobulins, immunoglobulin chains orantibody fragments (Fv, Fab, Fab′, F(ab′)₂ or other antigen bindingsubsequences of antibodies) that contain minimal sequence derived fromnon-human immunoglobulin. In some embodiments, humanized antibodies arehuman immunoglobulin (recipient antibody) in which residues from acomplementary-determining region (CDR) of the recipient are replaced byresidues from a CDR of a non-human species (donor antibody) such asmouse, rat or rabbit having the desired specificity, affinity andcapacity. In particular embodiments, antibodies for use in the presentinvention bind to particular epitopes of CD20. In some embodiments,epitopes of CD20 to which anti-CD20 antibodies bind include, forexample, ¹⁷⁰ANPS¹⁷³ (Binder et al., Blood 2006, 108(6): 1975-1978), FMC7(Deans et al., Blood 2008, 111(4): 2492), Rp5-L and Rp15-C (mimotopes ofCD20) (Perosa et al., J. Immunol. 2009, 182:416-423), ¹⁸²YCYSI¹⁸⁵(Binder et al., Blood 2006, 108(6): 1975-1978) and WEWTI (a mimic of¹⁸²YCYSI¹⁸⁵) (Binder et al., Blood 2006, 108(6): 1975-1978). In someembodiments, an anti-CD20 antibody has a binding affinity (K_(d)) for anepitope of CD20 of less than 12 nM, less than 11 nM, less than 10 nM,less than 9 nM, less than 8 nM, less than 7 nM, less than 6 nM, lessthan 5 nM, less than 4 nM, less than 3 nM, less than 2 nM or less than 1nM.

As used herein, the term “biosimilar” (for example, of an approvedreference product/biological drug, such as a protein therapeutic,antibody, etc.) refers to a biologic product that is similar to thereference product based upon data derived from (a) analytical studiesthat demonstrate that the biological product is highly similar to thereference product notwithstanding minor differences in clinicallyinactive components; (b) animal studies (including the assessment oftoxicity); and/or (c) a clinical study or studies (including theassessment of immunogenicity and pharmacokinetics or pharmacodynamics)that are sufficient to demonstrate safety, purity, and potency in one ormore appropriate conditions of use for which the reference product isapproved and intended to be used and for which approval is sought (e.g.,that there are no clinically meaningful differences between thebiological product and the reference product in terms of the safety,purity, and potency of the product).

In some embodiments, the biosimilar biological product and referenceproduct utilizes the same mechanism or mechanisms of action for thecondition or conditions of use prescribed, recommended, or suggested inthe proposed labeling, but only to the extent the mechanism ormechanisms of action are known for the reference product. In someembodiments, the condition or conditions of use prescribed, recommended,or suggested in the labeling proposed for the biological product havebeen previously approved for the reference product. In some embodiments,the route of administration, the dosage form, and/or the strength of thebiological product are the same as those of the reference product. Insome embodiments, the facility in which the biological product ismanufactured, processed, packed, or held meets standards designed toassure that the biological product continues to be safe, pure, andpotent. The reference product may be approved in at least one of theU.S., Europe, or Japan. A biosimilar can be for example, a presentlyknown antibody having the same primary amino acid sequence as a marketedantibody, but may be made in different cell types or by differentproduction, purification or formulation methods.

As used herein, the terms “combination”, “in combination with” or“combination therapy” refer to those situations in which two or moredifferent pharmaceutical agents are administered in overlapping regimensso that the subject is simultaneously exposed to both agents. In someembodiments, such combinations refer to simultaneously administering toa subject separate dosage forms of Compound 1 and rituximab. In someembodiments, such combinations refer to contemporaneously administeringto a subject separate dosage forms of Compound 1 and rituximab, whereinCompound 1 is administered before, during or after administration ofrituximab. In some embodiments, simultaneous or contemporaneous exposureof Compound 1 and rituximab is effected via different dosage regimensappropriate for each therapeutic agent. For example, Compound 1 may beadministered once or twice daily for one or more 28-day cycles, whereasrituximab may be administered once during a 28-day cycle.

The term “percent inhibition” as used herein refers to the percentdecrease of target activity in the presence of a test compound (e.g., anirreversible BTK inhibitor) relative to control target activity. It willbe appreciated that percent inhibition of a target (e.g., a kinase) canbe determined in numerous ways, one of which is described in Example 2,infra. In some embodiments, percent inhibition is expressed as %inhibition (e.g., 50% inhibition). In some embodiments, the percentinhibition of a kinase is an average percent inhibition.

As used herein, the term “comparable”, refers to two or more agents,entities, situations, sets of conditions, etc. that may not be identicalto one another but that are sufficiently similar to permit comparisontherebetween so that conclusions may reasonably be drawn based ondifferences or similarities observed. Those of ordinary skill in the artwill understand, in context, what degree of identity is required in anygiven circumstance for two or more such agents, entities, situations,sets of conditions, etc. to be considered comparable. As used herein,the terms “comparable percent inhibition” or “comparable average percentinhibition” refer to a percent inhibition or an average percentinhibition, respectively, of a kinase that is within 10% of thatobserved or determined for a reference kinase inhibitor. For example, ifa reference kinase inhibitor has 50% inhibition of a kinase relative toa control, another inhibitor will be considered to show comparableinhibition if it has about 40% to about 60% inhibition of the samekinase relative to the control. In some embodiments, an irreversible BTKinhibitor has comparable percent inhibition to a reference kinaseinhibitor wherein the percent inhibition of the irreversible BTKinhibitor is within 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1% inhibition ofthat observed or determined for a reference kinase inhibitor.

As used herein, a “disease or disorder associated with BTK” or a“BTK-mediated disorder” means any disease or other deleterious conditionin which BTK, or a mutant thereof, is known or suspected to play a role.Accordingly, another embodiment of the present invention relates topreventing, treating, stabilizing or lessening the severity orprogression of one or more diseases in which BTK, or a mutant thereof,is known or suspected to play a role. Specifically, the presentinvention relates to a method of treating or lessening the severity of aproliferative disorder, wherein said method comprises administering to apatient in need thereof Compound 1 in combination with rituximab.

As used herein, the term “irreversible” or “irreversible inhibitor”refers to an inhibitor (i.e. a compound) that is able to be covalentlybonded to a target protein kinase in a substantially non-reversiblemanner. That is, whereas a reversible inhibitor is able to bind to (butis generally unable to form a covalent bond) the target protein kinase,and therefore can become dissociated from the target protein kinase, anirreversible inhibitor will remain substantially bound to the targetprotein kinase once covalent bond formation has occurred. Irreversibleinhibitors usually display time dependency, whereby the degree ofinhibition increases with the time with which the inhibitor is incontact with the enzyme. Methods for identifying if a compound is actingas an irreversible inhibitor are known to one of ordinary skill in theart. Such methods include, but are not limited to, enzyme kineticanalysis of the inhibition profile of the compound with the proteinkinase target, the use of mass spectrometry of the protein drug targetmodified in the presence of the inhibitor compound, discontinuousexposure, also known as “washout,” experiments, and the use of labeling,such as radiolabelled inhibitor, to show covalent modification of theenzyme, as well as other methods known to one of skill in the art.

The term “refractory CLL/SLL” as used herein is defined as CLL/SLL whichwas treated with at least one line of prior therapy (i) withoutachieving at least a partial response to therapy or (ii) whichprogressed within 6 months of treatment.

The term “relapsed CLL/SLL” as used herein is defined as CLL/SLL whichprogressed after ≧6 months post-treatment after achieving partialresponse or complete response to therapy.

The term “subject”, as used herein, means a mammal and includes humanand animal subjects, such as domestic animals (e.g., horses, dogs, cats,etc.).

As used herein, a “therapeutically effective amount” means an amount ofa substance (e.g., a therapeutic agent, composition, and/or formulation)that elicits a desired biological response. In some embodiments, atherapeutically effective amount of a substance is an amount that issufficient, when administered as part of a dosing regimen to a subjectsuffering from or susceptible to a disease, disorder, and/or condition,to treat, diagnose, prevent, and/or delay the onset of the disease,disorder, and/or condition. As will be appreciated by those of ordinaryskill in this art, the effective amount of a substance may varydepending on such factors as the desired biological endpoint, thesubstance to be delivered, the target cell or tissue, etc. For example,the effective amount of compound in a formulation to treat a disease,disorder, and/or condition is the amount that alleviates, ameliorates,relieves, inhibits, prevents, delays onset of, reduces severity ofand/or reduces incidence of one or more symptoms or features of thedisease, disorder, and/or condition. In some embodiments, a“therapeutically effective amount” is at least a minimal amount of acompound, or composition containing a compound, which is sufficient fortreating one or more symptoms of a disorder or condition associated withBruton's tyrosine kinase.

The terms “treat” or “treating,” as used herein, refers to partially orcompletely alleviating, inhibiting, delaying onset of, preventing,ameliorating and/or relieving a disorder or condition, or one or moresymptoms of the disorder or condition. As used herein, the terms“treatment,” “treat,” and “treating” refer to partially or completelyalleviating, inhibiting, delaying onset of, preventing, amelioratingand/or relieving a disorder or condition, or one or more symptoms of thedisorder or condition, as described herein. In some embodiments,treatment may be administered after one or more symptoms have developed.In some embodiments, the term “treating” includes preventing or haltingthe progression of a disease or disorder. In other embodiments,treatment may be administered in the absence of symptoms. For example,treatment may be administered to a susceptible individual prior to theonset of symptoms (e.g., in light of a history of symptoms and/or inlight of genetic or other susceptibility factors). Treatment may also becontinued after symptoms have resolved, for example to prevent or delaytheir recurrence. Thus, in some embodiments, the term “treating”includes preventing relapse or recurrence of a disease or disorder.

The expression “unit dosage form” as used herein refers to a physicallydiscrete unit of therapeutic formulation appropriate for the subject tobe treated. It will be understood, however, that the total daily usageof the compositions of the present invention will be decided by theattending physician within the scope of sound medical judgment. Thespecific effective dose level for any particular subject or organismwill depend upon a variety of factors including the disorder beingtreated and the severity of the disorder; activity of specific activeagent employed; specific composition employed; age, body weight, generalhealth, sex and diet of the subject; time of administration, and rate ofexcretion of the specific active agent employed; duration of thetreatment; drugs and/or additional therapies used in combination orcoincidental with specific compound(s) employed, and like factors wellknown in the medical arts.

General Methods of Treating a BTK-Mediated Disease or Disorder

In some embodiments, the present invention provides a method oftreating, stabilizing or lessening the severity or progression of one ormore diseases and conditions associated with BTK comprisingadministering to a patient in need thereof an irreversible BTK inhibitorand rituximab. In some such embodiments, provided methods furthercomprise administering fludarabine and cyclophosphamide.

It is understood that although the methods described herein refer toformulations, doses and dosing regimens/schedules of Compound 1 andsalts thereof, such formulations, doses and/or dosing regimens/schedulesare equally applicable to any irreversible BTK inhibitor, such as thosedescribed below. Accordingly, in some embodiments, a dose or dosingregimen of an irreversible BTK inhibitor is selected from any of thedoses or dosing regimens for Compound 1 as described herein. In someembodiments, provided methods comprise administering an irreversible BTKinhibitor in an amount selected from any of the doses for Compound 1 asdescribed herein. In some such embodiments, a dose of an irreversibleBTK inhibitor is administered according to a dosing schedule selectedfrom any of the dosing schedules described herein for Compound 1. Insome embodiments, a composition comprising an irreversible BTK inhibitoris any of the formulations as described herein.

In some embodiments, the present invention provides a method oftreating, stabilizing or lessening the severity or progression of one ormore diseases and conditions associated with BTK comprisingadministering to a patient in need thereof an irreversible BTK inhibitorand rituximab and bendamustine.

In some embodiments, the irreversible BTK inhibitor covalently binds toCys 481 of BTK.

In some embodiments, an irreversible BTK inhibitor has activity againstone or more kinases selected from the kinases recited in Table 3, infra.

In some embodiments, an irreversible BTK inhibitor has a percentinhibition comparable to that of a reference kinase inhibitor withrespect to a kinase selected from Table 3, or combinations thereof. Insome such embodiments, the reference kinase inhibitor is Compound 2:

In some embodiments, the percent inhibition of the reference kinaseinhibitor is that shown for Compound 2 in Example 2.

In some embodiments, an irreversible BTK inhibitor has a percentinhibition comparable to that of Compound 2 with respect to one or morekinases selected from Table 3, or combinations thereof, in that theirreversible kinase inhibitor has a percent inhibition withinapproximately 10% of that observed for Compound 2. In some embodiments,an irreversible BTK inhibitor has a percent inhibition comparable tothat of Compound 2 with respect to one or more kinases selected fromTable 3, or combinations thereof, in that the irreversible kinaseinhibitor has a percent inhibition that is within about 9%, or about 8%,or about 7%, or about 6%, or about 5%, or about 4%, or about 3%, orabout 2% or about 1% inhibition of that observed for Compound 2.

In some embodiments, an irreversible BTK inhibitor has a percentinhibition that is greater than that observed for Compound 2 withrespect to one or more kinases selected from Table 3. In someembodiments, an irreversible BTK inhibitor has a percent inhibition thatis less than that observed for Compound 2 with respect to one or morekinases selected from Table 3.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition of one or more additional kinases, wherein the percentinhibition of the kinase or kinases is at least about 50%, at leastabout 55%, at least about 60%, at least about 65%, at least about 70%,at least about 75%, at least about 80%, at least about 85%, at leastabout 90% or at least about 95%.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition comparable to that of a reference kinase inhibitor withrespect to one or more kinases selected from the group consisting ofTXK, BMX/ETK, FLT3, BLK, TEC, ERBB4/HER4, Aurora B, TRKC, RET,LOK/STK10, Aurora C, FLT4/VEGFR3, ROS/ROS1, ARK5/NUAK1, EGFR, DDR1,JAK3, LRRK2, ABL2/ARG, ITK, Aurora A, YES/YES1, FGFR3, TNK1, BRK, FGFR2,PDGFRb, c-SRC, ACK1, FGFR1, STK16, ABL1, AXL, TYK2, ERBB2/HER2, FGR,CHK2, SIK1, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, PKCb2 and CLK2, orcombinations thereof. In some embodiments, the reference kinaseinhibitor is Compound 2. In some embodiments, the percent inhibition ofthe reference kinase inhibitor is that shown for Compound 2 in Example2.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition comparable to that of a reference kinase inhibitor withrespect to the group of kinases consisting of TXK, BMX/ETK, FLT3, BLK,TEC, ERBB4/HER4, Aurora B, TRKC, RET, LOK/STK10, Aurora C, FLT4/VEGFR3,ROS/ROS1, ARK5/NUAK1, EGFR, DDR1, JAK3, LRRK2, ABL2/ARG, ITK, Aurora A,YES/YES1, FGFR3, TNK1, BRK, FGFR2, PDGFRb, c-SRC, ACK1, FGFR1, STK16,ABL1, AXL, TYK2, ERBB2/HER2, FGR, CHK2, SIK1, MLK1/MAP3K9, MLK2/MAP3K10,MLK3/MAP3K11, PKCb2 and CLK2, or combinations thereof. In someembodiments, the reference kinase inhibitor is Compound 2. In someembodiments, the percent inhibition of the reference kinase inhibitor isthat shown for Compound 2 in Example 2.

In some embodiments, the irreversible BTK inhibitor inhibits a kinaseselected from the group consisting of TXK, BMX/ETK, FLT3, BLK, TEC,ERBB4/HER4, Aurora B, TRKC, RET, LOK/STK10, Aurora C, FLT4/VEGFR3,ROS/ROS1, ARK5/NUAK1, EGFR, DDR1, JAK3, LRRK2, ABL2/ARG, ITK, Aurora A,YES/YES1, FGFR3, TNK1, BRK, FGFR2, PDGFRb, c-SRC, ACK1, FGFR1, STK16,ABL1, AXL, TYK2, ERBB2/HER2, FGR, CHK2, SIK1, MLK1/MAP3K9, MLK2/MAP3K10,MLK3/MAP3K11, PKCb2 and CLK2, or combinations thereof, wherein theinhibition of the kinase or kinases is at least the percent inhibitionobserved for a reference kinase inhibitor. In some embodiments, thereference kinase inhibitor is Compound 2. In some embodiments, thepercent inhibition of the reference kinase inhibitor is that shown forCompound 2 in Example 2.

In some embodiments, the irreversible BTK inhibitor has at least about50%, at least about 55%, at least about 60%, at least 65%, at leastabout 70%, at least about 75%, at least about 80%, at least about 85%,at least about 90% or at least about 95% inhibition of a kinase selectedfrom TXK, BMX/ETK, FLT3, BLK, TEC, ERBB4/HER4, Aurora B, TRKC, RET,LOK/STK10, Aurora C, FLT4/VEGFR3, ROS/ROS1, ARK5/NUAK1, EGFR, DDR1,JAK3, LRRK2, ABL2/ARG, ITK, Aurora A, YES/YES1, FGFR3, TNK1, BRK, FGFR2,PDGFRb, c-SRC, ACK1, FGFR1, STK16, ABL1, AXL, TYK2, ERBB2/HER2, FGR,CHK2, SIK1, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, PKCb2 and CLK2, orcombinations thereof.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition comparable to that of a reference kinase inhibitor withrespect to one or more kinases selected from the group consisting ofAurora A, Aurora B, Aurora C, TRKC, ROS/ROS1, ARK5/NUAK1, LRRK2,ABL2/ARG, TNK1, STK16, ABL1, AXL, TYK2, CHK2, SIK1, MLK1/MAP3K9,MLK2/MAP3K10, MLK3/MAP3K11, PKCb2 and CLK2, or combinations thereof. Insome embodiments, the reference kinase inhibitor is Compound 2. In someembodiments, the percent inhibition of the reference kinase inhibitor isthat shown for Compound 2 in Example 2.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition comparable to that of a reference kinase inhibitor withrespect to the group of kinases consisting of Aurora A, Aurora B, AuroraC, TRKC, ROS/ROS1, ARK5/NUAK1, LRRK2, ABL2/ARG, TNK1, STK16, ABL1, AXL,TYK2, CHK2, SIK1, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, PKCb2 andCLK2, or combinations thereof. In some embodiments, the reference kinaseinhibitor is Compound 2. In some embodiments, the percent inhibition ofthe reference kinase inhibitor is that shown for Compound 2 in Example2.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition that is at least the percent inhibition observed for areference kinase inhibitor with respect to one or more kinases selectedfrom the group consisting of Aurora A, Aurora B, Aurora C, TRKC,ROS/ROS1, ARK5/NUAK1, LRRK2, ABL2/ARG, TNK1, STK16, ABL1, AXL, TYK2,CHK2, SIK1, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, PKCb2 and CLK2, orcombinations thereof. In some embodiments, the reference kinaseinhibitor is Compound 2. In some embodiments, the percent inhibition ofthe reference kinase inhibitor is that shown for Compound 2 in Example2.

In some embodiments, the irreversible BTK inhibitor has at least about50%, at least about 55%, at least about 60%, at least about 65%, atleast about 70%, at least about 75%, at least about 80%, at least about85%, at least about 90% or at least about 95% inhibition of a kinaseselected from Aurora A, Aurora B, Aurora C, TRKC, ROS/ROS1, ARK5/NUAK1,LRRK2, ABL2/ARG, TNK1, STK16, ABL1, AXL, TYK2, CHK2, SIK1, MLK1/MAP3K9,MLK2/MAP3K10, MLK3/MAP3K11, PKCb2 and CLK2, or combinations thereof.

In some embodiments, the present invention provides a method oftreating, stabilizing or lessening the severity or progression ofCLL/SLL comprising administering to a patient in need thereof anirreversible BTK inhibitor in combination with rituximab, wherein theirreversible BTK inhibitor has not more than about 50% inhibition of akinase selected from Aurora A, Aurora B, Aurora C, TRKC, ROS/ROS1,ARK5/NUAK1, LRRK2, ABL2/ARG, TNK1, STK16, ABL1, AXL, TYK2, CHK2, SIK1,MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, PKCb2 and CLK2, or combinationsthereof.

In some embodiments, the irreversible BTK inhibitor has at least about50% inhibition of a kinase selected from Aurora A, Aurora B, Aurora C,TRKC, ROS/ROS1, ARK5/NUAK1, LRRK2, ABL2/ARG, TNK1, STK16, ABL1, AXL,TYK2, CHK2, SIK1, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11 and PKCb2, orcombinations thereof.

In some embodiments, the irreversible BTK inhibitor has has at leastabout 50% inhibition of the group of kinases consisting of Aurora A,Aurora B, Aurora C, TRKC, ROS/ROS1, ARK5/NUAK1, LRRK2, ABL2/ARG, TNK1,STK16, ABL1, AXL, TYK2, CHK2, SIK1, MLK1/MAP3K9, MLK2/MAP3K10,MLK3/MAP3K11, PKCb2 and CLK2.

In some embodiments, the irreversible BTK inhibitor has at least about55% inhibition of a kinase selected from Aurora A, Aurora B, Aurora C,TRKC, ROS/ROS1, ARK5/NUAK1, LRRK2, ABL2/ARG, TNK1, STK16, ABL1, CHK2,MLK1/MAP3K9, MLK2/MAP3K10 and MLK3/MAP3K11, or combinations thereof. Insome embodiments, the irreversible BTK inhibitor has at least about 55%inhibition of the group of kinases consisting of Aurora A, Aurora B,Aurora C, TRKC, ROS/ROS1, ARK5/NUAK1, LRRK2, ABL2/ARG, TNK1, STK16,ABL1, CHK2, MLK1/MAP3K9, MLK2/MAP3K10 and MLK3/MAP3K11.

In some embodiments, the irreversible BTK inhibitor has at least about60% inhibition of a kinase selected from Aurora A, Aurora B, Aurora C,TRKC, ROS/ROS1, ARK5/NUAK1, LRRK2, ABL2/ARG, TNK1, STK16, CHK2,MLK1/MAP3K9 and MLK3/MAP3K11, or combinations thereof. In someembodiments, the irreversible BTK inhibitor has at least about 60%inhibition of the group of kinases consisting of Aurora A, Aurora B,Aurora C, TRKC, ROS/ROS1, ARK5/NUAK1, LRRK2, ABL2/ARG, TNK1, STK16,CHK2, MLK1/MAP3K9 and MLK3/MAP3K11.

In some embodiments, the irreversible BTK inhibitor has at least about65% inhibition of a kinase selected from Aurora A, Aurora B, Aurora C,TRKC, ROS/ROS1, ARK5/NUAK1, LRRK2, TNK1, STK16, CHK2, MLK1/MAP3K9 andMLK3/MAP3K11, or combinations thereof. In some embodiments, theirreversible BTK inhibitor sh at least about 65% inhibition of the groupof kinases consisting of Aurora A, Aurora B, Aurora C, TRKC, ROS/ROS1,ARK5/NUAK1, LRRK2, TNK1, STK16, CHK2, MLK1/MAP3K9 and MLK3/MAP3K11.

In some embodiments, the irreversible BTK inhibitor has at least about70% inhibition of a kinase selected from Aurora A, Aurora B, Aurora C,ROS/ROS1, ARK5/NUAK1, TNK1, STK16, CHK2, MLK1/MAP3K9 and MLK3/MAP3K11,or combinations thereof. In some embodiments, the irreversible BTKinhibitor at least about 70% inhibition of the group of kinasesconsisting of Aurora A, Aurora B, Aurora C, ROS/ROS1, ARK5/NUAK1, TNK1,STK16, CHK2, MLK1/MAP3K9 and MLK3/MAP3K11.

In some embodiments, the irreversible BTK inhibitor has at least about75% inhibition of a kinase selected from Aurora A, Aurora B, ROS/ROS1,ARK5/NUAK1, TNK1, STK16 and MLK1/MAP3K9, or combinations thereof. Insome embodiments, the irreversible BTK inhibitor has at least about 75%inhibition of the group of kinases consisting of Aurora A, Aurora B,ROS/ROS1, ARK5/NUAK1, TNK1, STK16 and MLK1/MAP3K9.

In some embodiments, the irreversible BTK inhibitor has at least about80% inhibition of a kinase selected from Aurora A, Aurora B, ROS/ROS1,ARK5/NUAK1, TNK1 and MLK1/MAP3K9, or combinations thereof. In someembodiments, the irreversible BTK inhibitor has at least about 80%inhibition of the group of kinases consisting of Aurora A, Aurora B,ROS/ROS1, ARK5/NUAK1, TNK1 and MLK1/MAP3K9.

In some embodiments, the irreversible BTK inhibitor has at least about85% inhibition of a kinase selected from Aurora A, Aurora B, ROS/ROS1,ARK5/NUAK1 and MLK1/MAP3K9, or combinations thereof. In someembodiments, the irreversible BTK inhibitor has at least about 85%inhibition of the group of kinases consisting of Aurora A, Aurora B,ROS/ROS1, ARK5/NUAK1 and MLK1/MAP3K9.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition comparable to that of a reference kinase inhibitor withrespect to one or more kinases selected from the group consisting ofTNK1, STK16, ABL1, AXL, TYK2, CHK2, MLK1/MAP3K9, MLK2/MAP3K10,MLK3/MAP3K11, SIK1, PKCb2 and CLK2, or combinations thereof. In someembodiments, the reference kinase inhibitor is Compound 2. In someembodiments, the percent inhibition of the reference kinase inhibitor isthat shown for Compound 2 in Example 2.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition comparable to that of a reference kinase inhibitor withrespect to the group of kinases consisting of TNK1, STK16, ABL1, AXL,TYK2, CHK2, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, SIK1, PKCb2 andCLK2, or combinations thereof. In some embodiments, the reference kinaseinhibitor is Compound 2. In some embodiments, the percent inhibition ofthe reference kinase inhibitor is that shown for Compound 2 in Example2.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition that is at least the percent inhibition observed for areference kinase inhibitor with respect to one or more kinases selectedfrom the group consisting of TNK1, STK16, ABL1, AXL, TYK2, CHK2,MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, SIK1, PKCb2 and CLK2, orcombinations thereof. In some embodiments, the reference kinaseinhibitor is Compound 2. In some embodiments, the percent inhibition ofthe reference kinase inhibitor is that shown for Compound 2 in Example2.

In some embodiments, the irreversible BTK inhibitor has at least about50%, at least about 55%, at least about 60%, at least about 65%, atleast about 70%, at least about 75%, at least about 80%, at least about85%, at least about 90% or at least about 95% inhibition of a kinaseselected from TNK1, STK16, ABL1, AXL, TYK2, CHK2, MLK1/MAP3K9,MLK2/MAP3K10, MLK3/MAP3K11, SIK1, PKCb2 and CLK2, or combinationsthereof.

In some embodiments, the irreversible BTK inhibitor has at least about50% inhibition of a kinase selected from TNK1, STK16, ABL1, AXL, TYK2,CHK2, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11, SIK1, PKCb2 and CLK2, orcombinations thereof. In some embodiments, the irreversible BTKinhibitor at least about 50% inhibition of the group of kinasesconsisting of TNK1, STK16, ABL1, AXL, TYK2, CHK2, MLK1/MAP3K9,MLK2/MAP3K10, MLK3/MAP3K11, SIK1, PKCb2 and CLK2.

In some embodiments, the irreversible BTK inhibitor has at least about55% inhibition of a kinase selected from TNK1, STK16, ABL1, CHK2,MLK1/MAP3K9 and MLK3/MAP3K11, or combinations thereof. In someembodiments, the irreversible BTK inhibitor has at least about 55%inhibition of the group of kinases consisting of TNK1, STK16, ABL1,CHK2, MLK1/MAP3K9 and MLK3/MAP3K11, or combinations thereof.

In some embodiments, the irreversible BTK inhibitor has at least about60%, at least about 65% or at least about 70% inhibition of a kinaseselected from TNK1, STK16, CHK2, MLK1/MAP3K9 and MLK3/MAP3K11, orcombinations thereof. In some embodiments, the irreversible BTKinhibitor has at least about 60%, at least about 65% or at least about70% inhibition of the group of kinases consisting of CHK2, MLK1/MAP3K9and MLK3/MAP3K11.

In some embodiments, the irreversible BTK inhibitor has at least about75% inhibition of a kinase selected from TNK1, STK16 and MLK1/MAP3K9, orcombinations thereof. In some embodiments, the irreversible BTKinhibitor has at least about 75% inhibition of the the group of kinasesconsisting of TNK1, STK16 and MLK1/MAP3K9.

In some embodiments, an irreversible BTK inhibitor for use in thepresent invention has a percent inhibition comparable to that of areference kinase inhibitor with respect to one or more kinases selectedfrom the group consisting of c-Kit, PDGFRa, RIPK2, HCK, EPHA6, LYN, CSK,LCK, ZAK/MILTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS, orcombinations thereof. In some embodiments, the reference kinaseinhibitor is Compound 2. In some embodiments, the percent inhibition ofthe reference kinase inhibitor is that shown for Compound 2 in Example2.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition comparable to that of a reference kinase inhibitor withrespect to the group of kinases consisting of c-Kit, PDGFRa, RIPK2, HCK,EPHA6, LYN, CSK, LCK, ZAK/MILTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF, ARAFand SRMS, or combinations thereof. In some embodiments, the referencekinase inhibitor is Compound 2. In some embodiments, the percentinhibition of the reference kinase inhibitor is that shown for Compound2 in Example 2.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition that is not more than the percent inhibition observed for areference kinase inhibitor with respect to one or more kinases selectedfrom the group consisting of c-Kit, PDGFRa, RIPK2, HCK, EPHA6, LYN, CSK,LCK, ZAK/MILTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS, orcombinations thereof. In some embodiments, the reference kinaseinhibitor is Compound 2. In some embodiments, the percent inhibition ofthe reference kinase inhibitor is that shown for Compound 2 in Example2.

In some embodiments, the irreversible BTK inhibitor has not more thanabout 50%, not more than about 45%, not more than about 40%, not morethan about 35%, not more than about 30%, not more than about 25%, notmore than about 20%, not more than about 15%, not more than about 10% ornot more than about 5% inhibition of a kinase selected from c-Kit,PDGFRa, RIPK2, HCK, EPHA6, LYN, CSK, LCK, ZAK/MLTK, LYN B, FRK/PTK5,FYN, RIPK3, BRAF, ARAF and SRMS, or combinations thereof.

In some embodiments, the irreversible BTK inhibitor has not more thanabout 50%, not more than about 45%, not more than about 40%, not morethan about 35%, not more than about 30%, not more than about 25%, notmore than about 20%, not more than about 15%, not more than about 10% ornot more than about 5% inhibition of a kinase selected from RIPK2, HCK,LYN, CSK, LCK, LYN B and FYN, or combinations thereof.

In some embodiments, the irreversible BTK inhibitor has not more thanabout 50%, not more than about 45%, not more than about 40%, not morethan about 35%, not more than about 30%, not more than about 25%, notmore than about 20%, not more than about 15%, not more than about 10% ornot more than about 5% inhibition of a kinase selected from EPHA6, LYNB, FRK/PTK5, RIPK3, BRAF, ARAF and SRMS, or combinations thereof.

In some embodiments, the present invention provides a method oftreating, stabilizing or lessening the severity or progression ofCLL/SLL comprising administering to a patient in need thereof anirreversible BTK inhibitor in combination with rituximab, wherein theirreversible BTK inhibitor has at least about 50% inhibition of a kinaseselected from c-Kit, PDGFRa, RIPK2, HCK, EPHA6, LYN, CSK, LCK, ZAK/MLTK,LYN B, FRK/PTK5, FYN, BRAF, RIPK3, ARAF and SRMS, or combinationsthereof.

In some embodiments, the irreversible BTK inhibitor has not more thanabout 50% inhibition of a kinase selected from c-Kit, PDGFRa, RIPK2,HCK, EPHA6, LYN, CSK, LCK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF,ARAF and SRMS, or combinations thereof. In some embodiments, theirreversible BTK inhibitor has not more than about 50% inhibition of thegroup of kinases consisting of c-Kit, PDGFRa, RIPK2, HCK, EPHA6, LYN,CSK, LCK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS.

In some embodiments, the irreversible BTK inhibitor has not more thanabout 45% inhibition of a kinase selected from c-Kit, PDGFRa, RIPK2,HCK, EPHA6, LYN, CSK, LCK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF,ARAF and SRMS, or combinations thereof. In some embodiments, theirreversible BTK inhibitor has not more than about 45% inhibition of thegroup of kinases consisting of c-Kit, PDGFRa, RIPK2, HCK, EPHA6, LYN,CSK, LCK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS.

In some embodiments, the irreversible BTK inhibitor has not more thanabout 40% inhibition of a kinase selected from c-Kit, PDGFRa, RIPK2,HCK, EPHA6, LYN, CSK, LCK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF,ARAF and SRMS, or combinations thereof. In some embodiments, theirreversible BTK inhibitor has not more than about 40% inhibition of thegroup of kinases consisting of c-Kit, PDGFRa, RIPK2, HCK, EPHA6, LYN,CSK, LCK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS.

In some embodiments, the irreversible BTK inhibitor has not more thanabout 35% inhibition of a kinase selected from c-Kit, RIPK2, HCK, EPHA6,LYN, CSK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS, orcombinations thereof. In some embodiments, the irreversible BTKinhibitor has not more than about 35% inhibition of the group of kinasesconsisting of c-Kit, RIPK2, HCK, EPHA6, LYN, CSK, ZAK/MLTK, LYN B,FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS.

In some embodiments, the irreversible BTK inhibitor has not more thanabout 30% inhibition of a kinase selected from c-Kit, RIPK2, HCK, EPHA6,LYN, CSK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS, orcombinations thereof. In some embodiments, the irreversible BTKinhibitor has not more than about 30% inhibition of the group of kinasesconsisting of c-Kit, RIPK2, HCK, EPHA6, LYN, CSK, ZAK/MLTK, LYN B,FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS.

In some embodiments, the irreversible BTK inhibitor has not more thanabout 25% inhibition of a kinase selected from c-Kit, RIPK2, EPHA6, CSK,ZAK/MLTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS, orcombinations thereof. In some embodiments, the irreversible BTKinhibitor has not more than about 25% inhibition of the group of kinasesconsisting of c-Kit, IPK2, EPHA6, CSK, ZAK/MLTK, LYN B, FRK/PTK5, FYN,RIPK3, BRAF, ARAF and SRMS.

In some embodiments, the irreversible BTK inhibitor has not more thanabout 20% inhibition of a kinase selected from EPHA6, CSK, ZAK/MLTK, LYNB, FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS, or combinations thereof.In some embodiments, the irreversible BTK inhibitor has not more thanabout 20% inhibition of the group of kinases consisting of EPHA6, CSK,ZAK/MLTK, LYN B, FRK/PTK5, FYN, RIPK3, BRAF, ARAF and SRMS.

In some embodiments, the irreversible BTK inhibitor has not more thanabout 15% inhibition of a kinase selected from EPHA6, LYN B, FRK/PTK5,RIPK3, BRAF, ARAF and SRMS, or combinations thereof. In someembodiments, the irreversible BTK inhibitor has not more than about 15%inhibition of the group of kinases consisting of EPHA6, LYN B, FRK/PTK5,RIPK3, BRAF, ARAF and SRMS.

In some embodiments, the irreversible BTK inhibitor has not more thanabout 10% inhibition of a kinase selected from EPHA6, LYN B, FRK/PTK5,RIPK3, BRAF, ARAF and SRMS, or combinations thereof. In someembodiments, the irreversible BTK inhibitor has not more than about 10%inhibition of the group of kinases consisting of EPHA6, LYN B, FRK/PTK5,RIPK3, BRAF, ARAF and SRMS.

In some embodiments, the irreversible BTK inhibitor has not more thanabout 5% inhibition of a kinase selected from EPHA6, FRK/PTK5, RIPK3,BRAF, ARAF and SRMS, or combinations thereof. In some embodiments, theirreversible BTK inhibitor has not more than about 5% inhibition of thegroup of kinases consisting of EPHA6, FRK/PTK5, RIPK3, BRAF, ARAF andSRMS.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition of LYN comparable to that of a reference kinase inhibitor. Insome embodiments, the irreversible BTK inhibitor has a percentinhibition of LYN that is not more than that observed for a referencekinase inhibitor. In some embodiments, the irreversible BTK inhibitorhas a percent inhibition of LYN that is about 20-30%. In someembodiments, the irreversible BTK inhibitor has a percent inhibition ofLYN that is about 25-30%. In some embodiments, the irreversible BTKinhibitor has a percent inhibition of LYN that is about 25-28%. In someembodiments, the irreversible BTK inhibitor has a percent inhibition ofLYN that is not more than about 25%, not more than about 26%, not morethan about 27%, not more than about 28%, not more than about 29%, notmore than about 30%, not more than about 31%, not more than about 32% ornot more than about 33%. In some embodiments, the reference kinaseinhibitor is Compound 2. In some embodiments, the percent inhibition ofthe reference kinase inhibitor is that shown for Compound 2 in Example2.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition of c-Kit comparable to that of a reference kinase inhibitor.In some embodiments, the irreversible BTK inhibitor has a percentinhibition of c-Kit that is not more than that observed for a referencekinase inhibitor. In some embodiments, the irreversible BTK inhibitorhas a percent inhibition of c-Kit that is about 15-25%. In someembodiments, the irreversible BTK inhibitor has a percent inhibition ofc-Kit that is about 20-25%. In some embodiments, the irreversible BTKinhibitor has a percent inhibition of c-Kit that is about 20-23%. Insome embodiments, the irreversible BTK inhibitor has a percentinhibition of c-Kit that is not more than about 15%, not more than about16%, not more than about 17%, not more than about 18%, not more thanabout 19%, not more than about 20%, not more than about 21%, not morethan about 22%, not more than about 23%, not more than about 24% or notmore than about 25%. In some embodiments, the reference kinase inhibitoris Compound 2. In some embodiments, the percent inhibition of thereference kinase inhibitor is that shown for Compound 2 in Example 2.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition of PDGFRa comparable to that of a reference kinase inhibitor.In some embodiments, the irreversible BTK inhibitor has a percentinhibition of PDGFRa that is not more than that observed for a referencekinase inhibitor. In some embodiments, the irreversible BTK inhibitorhas a percent inhibition of PDGFRa that is about 30-40%. In someembodiments, the irreversible BTK inhibitor has a percent inhibition ofPDGFRa that is about 35-40%. In some embodiments, the irreversible BTKinhibitor has a percent inhibition of PDGFRa that is about 35-38%. Insome embodiments, the irreversible BTK inhibitor has a percentinhibition of PDGFRa that is not more than about 30%, not more thanabout 31%, not more than about 32%, not more than about 33%, not morethan about 34%, not more than about 35%, not more than about 36%, notmore than about 37%, not more than about 38%, not more than about 39% ornot more than about 40%. In some embodiments, the reference kinaseinhibitor is Compound 2. In some embodiments, the percent inhibition ofthe reference kinase inhibitor is that shown for Compound 2 in Example2.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition of RIPK2 comparable to that of a reference kinase inhibitor.In some embodiments, the irreversible BTK inhibitor has a percentinhibition of RIPK2 that is not more than that observed for a referencekinase inhibitor. In some embodiments, the irreversible BTK inhibitorhas a percent inhibition of RIPK2 that is about 20-30%. In someembodiments, the irreversible BTK inhibitor has a percent inhibition ofRIPK2 that is about 20-25%. In some embodiments, the irreversible BTKinhibitor has a percent inhibition of RIPK2 that is about 22-25%. Insome embodiments, the irreversible BTK inhibitor has a percentinhibition of RIPK2 that is not more than about 18%, not more than about19%, not more than about 20%, not more than about 21%, not more thanabout 22%, not more than about 23%, not more than about 24%, not morethan about 25%, not more than about 26% or not more than about 27%. Insome embodiments, the reference kinase inhibitor is Compound 2. In someembodiments, the percent inhibition of the reference kinase inhibitor isthat shown for Compound 2 in Example 2.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition of HCK comparable to that of a reference kinase inhibitor. Insome embodiments, the irreversible BTK inhibitor has a percentinhibition of HCK that is not more than that observed for a referencekinase inhibitor. In some embodiments, the irreversible BTK inhibitorhas a percent inhibition of HCK that is about 25-35%. In someembodiments, the irreversible BTK inhibitor has a percent inhibition ofHCK that is about 27-32%. In some embodiments, the irreversible BTKinhibitor has a percent inhibition of HCK that is about 28-31%. In someembodiments, the irreversible BTK inhibitor has a percent inhibition ofHCK that is not more than about 26%, not more than about 27%, not morethan about 28%, not more than about 29%, not more than about 30%, notmore than about 31%, not more than about 32%, not more than about 33% ornot more than about 34%. In some embodiments, the reference kinaseinhibitor is Compound 2. In some embodiments, the percent inhibition ofthe reference kinase inhibitor is that shown for Compound 2 in Example2.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition of EPHA6 comparable to that of a reference kinase inhibitor.In some embodiments, the irreversible BTK inhibitor has a percentinhibition of EPHA6 that is not more than that observed for a referencekinase inhibitor. In some embodiments, the irreversible BTK inhibitorhas a percent inhibition of EPHA6 that is about 0-10%. In someembodiments, the irreversible BTK inhibitor has a percent inhibition ofEPHA6 that is about 0-5%. In some embodiments, the irreversible BTKinhibitor has a percent inhibition of EPHA6 that is about 0-3%. In someembodiments, the irreversible BTK inhibitor has a percent inhibition ofEPHA6 that is not more than about 0.5%, not more than about 0.6%, notmore than about 0.7%, not more than about 0.8%, not more than about0.9%, not more than about 1%, not more than about 2%, not more thanabout 3% or not more than about 4%. In some embodiments, the referencekinase inhibitor is Compound 2. In some embodiments, the percentinhibition of the reference kinase inhibitor is that shown for Compound2 in Example 2.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition of CSK comparable to that of a reference kinase inhibitor. Insome embodiments, the irreversible BTK inhibitor has a percentinhibition of CSK that is not more than that observed for a referencekinase inhibitor. In some embodiments, the irreversible BTK inhibitorhas a percent inhibition of CSK that is about 10-20%. In someembodiments, the irreversible BTK inhibitor has a percent inhibition ofCSK that is about 15-20%. In some embodiments, the irreversible BTKinhibitor has a percent inhibition of CSK that is about 16-19%. In someembodiments, the irreversible BTK inhibitor has a percent inhibition ofCSK that is not more than about 15%, not more than about 16%, not morethan about 17%, not more than about 18%, not more than about 19%, notmore than about 20%, not more than about 21%, not more than about 22% ornot more than about 23%. In some embodiments, the reference kinaseinhibitor is Compound 2. In some embodiments, the percent inhibition ofthe reference kinase inhibitor is that shown for Compound 2 in Example2.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition of LCK comparable to that of a reference kinase inhibitor. Insome embodiments, the irreversible BTK inhibitor has a percentinhibition of LCK that is not more than that observed for a referencekinase inhibitor. In some embodiments, the irreversible BTK inhibitorhas a percent inhibition of LCK that is about 30-40%. In someembodiments, the irreversible BTK inhibitor has a percent inhibition ofLCK that is about 32-37%. In some embodiments, the irreversible BTKinhibitor has a percent inhibition of LCK that is about 34-37%. In someembodiments, the irreversible BTK inhibitor has a percent inhibition ofLCK that is not more than about 34%, not more than about 35%, not morethan about 36%, not more than about 37%, not more than about 38%, notmore than about 39%, not more than about 40%, not more than about 41% ornot more than about 42%. In some embodiments, the reference kinaseinhibitor is Compound 2. In some embodiments, the percent inhibition ofthe reference kinase inhibitor is that shown for Compound 2 in Example2.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition of ZAK/MLTK comparable to that of a reference kinaseinhibitor. In some embodiments, the irreversible BTK inhibitor has apercent inhibition of ZAK/MLTK that is not more than that observed for areference kinase inhibitor. In some embodiments, the irreversible BTKinhibitor has a percent inhibition of ZAK/MLTK that is about 10-20%. Insome embodiments, the irreversible BTK inhibitor has a percentinhibition of ZAK/MLTK that is about 12-17%. In some embodiments, theirreversible BTK inhibitor has a percent inhibition of ZAK/MLTK that isabout 14-17%. In some embodiments, the irreversible BTK inhibitor has apercent inhibition of ZAK/MLTK that is not more than about 12%, not morethan about 13%, not more than about 14%, not more than about 15%, notmore than about 16%, not more than about 17%, not more than about 18%,not more than about 19% or not more than about 20%. In some embodiments,the reference kinase inhibitor is Compound 2. In some embodiments, thepercent inhibition of the reference kinase inhibitor is that shown forCompound 2 in Example 2.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition of LYN B comparable to that of a reference kinase inhibitor.In some embodiments, the irreversible BTK inhibitor has a percentinhibition of LYN B that is not more than that observed for a referencekinase inhibitor. In some embodiments, the irreversible BTK inhibitorhas a percent inhibition of LYN B that is about 0-10%. In someembodiments, the irreversible BTK inhibitor has a percent inhibition ofLYN B that is about 3-8%. In some embodiments, the irreversible BTKinhibitor has a percent inhibition of LYN B that is about 4-7%. In someembodiments, the irreversible BTK inhibitor has a percent inhibition ofLYN B that is not more than about 1%, not more than about 2%, not morethan about 3%, not more than about 4%, not more than about 5%, not morethan about 6%, not more than about 7%, not more than about 8%, not morethan about 9% or not more than about 10%. In some embodiments, thereference kinase inhibitor is Compound 2. In some embodiments, thepercent inhibition of the reference kinase inhibitor is that shown forCompound 2 in Example 2.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition of FRK/PTK5 comparable to that of a reference kinaseinhibitor. In some embodiments, the irreversible BTK inhibitor has apercent inhibition of FRK/PTK5 that is not more than that observed for areference kinase inhibitor. In some embodiments, the irreversible BTKinhibitor has a percent inhibition of FRK/PTK5 that is about 0-10%. Insome embodiments, the irreversible BTK inhibitor has a percentinhibition of FRK/PTK5 that is about 0-5%. In some embodiments, theirreversible BTK inhibitor has a percent inhibition of FRK/PTK5 that isabout 0-3%. In some embodiments, the irreversible BTK inhibitor has apercent inhibition of FRK/PTK5 that is not more than about 0.5%, notmore than about 0.6%, not more than about 0.7%, not more than about0.8%, not more than about 0.9%, not more than about 1%, not more thanabout 1.5%, not more than about 2%, not more than about 3% or not morethan about 4%. In some embodiments, the reference kinase inhibitor isCompound 2. In some embodiments, the percent inhibition of the referencekinase inhibitor is that shown for Compound 2 in Example 2.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition of FYN comparable to that of a reference kinase inhibitor. Insome embodiments, the irreversible BTK inhibitor has a percentinhibition of FYN that is not more than that observed for a referencekinase inhibitor. In some embodiments, the irreversible BTK inhibitorhas a percent inhibition of FYN that is about 15-25%. In someembodiments, the irreversible BTK inhibitor has a percent inhibition ofFYN that is about 15-20%. In some embodiments, the irreversible BTKinhibitor has a percent inhibition of FYN that is about 17-20%. In someembodiments, the irreversible BTK inhibitor has a percent inhibition ofFYN that is not more than about 15%, not more than about 16%, not morethan about 17%, not more than about 18%, not more than about 19%, notmore than about 20%, not more than about 21%, not more than about 22% ornot more than about 23%. In some embodiments, the reference kinaseinhibitor is Compound 2. In some embodiments, the percent inhibition ofthe reference kinase inhibitor is that shown for Compound 2 in Example2.

In some embodiments, the irreversible BTK inhibitor has a percentinhibition of BRAF comparable to that of a reference kinase inhibitor.In some embodiments, the irreversible BTK inhibitor has a percentinhibition of BRAF that is not more than that observed for a referencekinase inhibitor. In some embodiments, the irreversible BTK inhibitorhas a percent inhibition of BRAF that is about 0-10%. In someembodiments, the irreversible BTK inhibitor has a percent inhibition ofBRAF that is about 0.1-5%. In some embodiments, the irreversible BTKinhibitor has a percent inhibition of BRAF that is about 0.2-3%. In someembodiments, the irreversible BTK inhibitor has a percent inhibition ofBRAF that is not more than about 0.1%, not more than about 0.2%, notmore than about 0.3%, not more than about 0.4%, not more than about0.5%, not more than about 0.6%, not more than about 0.7%, not more thanabout 0.8%, not more than about 0.9%, not more than about 1%, not morethan about 2%, not more than about 3%, not more than about 4% or notmore than about 5%. In some embodiments, the reference kinase inhibitoris Compound 2. In some embodiments, the percent inhibition of thereference kinase inhibitor is that shown for Compound 2 in Example 2.

Compound 1 is an Irreversible BTK Inhibitor

As described above, Bruton's tyrosine kinase (Btk) is a non-receptortyrosine kinase with restricted cellular expression largely limited toB-lymphocytes, monocytes, and mast cells or basophils. Btk is a criticalcomponent of the B-cell receptor (BCR) signaling network and is crucialfor B-cell development. Investigation has revealed that some B-cellmalignancies, including CLL/SLL, depend on BCR signaling, suggestingthat interruption of such signaling could be a promising therapeuticopportunity. Recently, clinical anti-tumor responses in various B-cellnon-Hodgkin's lymphoma (NHL) and CLL/SLL have been reported with agentsthat inhibit spleen tyrosine kinase (Syk) and Btk, both components ofthe BCR signaling pathway.

Compound 1 is active in a variety of assays and therapeutic modelsdemonstrating covalent, irreversible inhibition of BTK (in enzymatic andcellular assays). Compound 1 inhibits Btk activity by binding with highaffinity to the adenosine triphosphate (ATP) binding site of Btk andforming a targeted covalent bond with the Btk protein, providing rapid,complete, and prolonged inhibition of Btk activity, both in vitro and invivo.

Phosphorylation of the auto-phosphorylation site on Btk (Tyr223) and theBtk responsive site (Tyr1217) on PLCγ2 in Ramos cells, a human Burkittlymphoma cell line, was inhibited by Compound 1 with an effectiveconcentration required for 50% inhibition (EC₅₀) of 1 nM to 10 nM.Compound 1 demonstrates a high degree of selectivity in cellular assaysystems against related kinases.

In single dose studies in healthy subjects, Compound 1 evidencedadequate safety, predictable pharmacokinetics (PK), and, at dosesgreater than 0.5 mg/kg, 80% to 100% occupancy of the Btk receptor targetin normal human peripheral blood B-cells. A phase I dose escalationstudy of a single agent of Compound 1 is currently being conducted indifferent hematologic malignancies, including CLL/SLL.

Anti-CD20 Antibodies

CD20, the first B-cell specific antigen defined by the monoclonalantibody tositumomab, plays a critical role in B-cell development. HumanCD20 is a 297 amino acid (30- to 35-kDa) phosphoprotein with fourtransmembrane domains encoded by the gene MS4A1 located on chromosome11q12.2. CD20 plays a critical role in B-cell development and is abiomarker for immunotherapies targeting B-cell derived diseases. CD20 isan integral membrane protein expressed by B lymphocytes in early stagesof differentiation and by most B cell lymphomas, but not bydifferentiated plasma cells. CD20 remains on the membrane of B cellswithout dissociation or internalization upon antibody binding. CD20functions though binding to the Src family of tyrosine kinases, such asLyn, Fyn and Lck, and believed to be involved as a result in thephosphorylation cascade of intracellular proteins. Anti-CD20 antibodiesare broadly classified into type I and type II antibodies. Both types ofanti-CD 20 antibodies exhibit equal ability in activating Fc-FcγRinteractions such as antibody-dependent cellular cytotoxicity (ADCC) andphagocytosis. Type I anti-CD20 antibodies redistribute CD20 intomembrane lipid rafts and potently activate complement-dependentcytotoxicity (CDC). Type II anti-CD20 antibodies weakly activate CDC butmore potently induce direct programmed cell death.

In some embodiments, the present invention encompasses the recognitionthat the combination of a BTK inhibitor, i.e. Compound 1, in combinationwith an anti-CD20 antibody is useful in treating BTK-mediated diseasesor disorders. Accordingly, in some embodiments, the present inventioncomprises a method of treating a BTK-mediated disease or disorder, themethod comprising administering to a patient in need thereof Compound 1in combination with an anti-CD20 antibody. A person of ordinary skill inthe art can readily identify and select additional anti-CD20 antibodiesthat are useful in the present invention. For example, in someembodiments, such antibodies are described, for example, in U.S. Pat.Nos. 8,153,125, 8,147,832, 8,101,179, 8,084,582, 8,057,793 and7,879,984, and U.S. Patent Publication Nos. 2011/0129412, 2012/0183545,2012/0134990 and 2012/0034185.

In some embodiments, an anti-CD20 antibody for use in the presentinvention is a type I antibody. In some embodiments, an anti-CD20 foruse in the present invention is a type II antibody.

In some embodiments, an anti-CD20 antibody is an antibody that binds toa CD20 epitope selected from ¹⁷⁰ANPS¹⁷³ and ¹⁸²CYSI¹⁸⁵.

In some embodiments, an anti-CD20 antibody has a binding affinity(K_(d)) for an epitope of CD20 of less than 12 nM, less than 11 nM, lessthan 10 nM, less than 9 nM, less than 8 nM, less than 7 nM, less than 6nM, less than 5 nM, less than 4 nM, less than 3 nM, less than 2 nM orless than 1 nM.

Rituximab is but one example of an anti-CD20 antibody. In someembodiments, an anti-CD20 antibody for use in the present inventionincludes, for example, rituximab (Rituxan® or MabThera®), Gazyva® (i.e.,obinutuzumab) and Arzerra® (ofatumumab). For ease of reference, providedmethods and regimens detailed herein refer to an exemplary anti-CD20antibody (i.e., rituximab); however, such reference is not intended tolimit the present invention to a single anti-CD20 antibody. Indeed, allreferences to rituximab, or a biosimilar thereof, are to be read by aperson skilled in the art to encompass the class of anti-CD20antibodies. For example, it will be appreciated that the anti-CD20antibodies ofatumumab (Arzerra®) or obinutuzumab (Gazyva®) can insteadbe administered in each instance where reference is made to rituximab.Thus, in some embodiments, provided methods comprise administeringCompound 1 and ofatumumab. In some such embodiments, ofatumumab isadministered in 12 doses according to the following schedule: 300 mginitial dose, followed 1 week later by 2000 mg dose weekly for 7 doses,followed 4 weeks later by 2000 mg every 4 weeks for 4 doses. In someembodiments, provided methods comprise administering Compound 1 andobinutuzumab. In some such embodiments, obinutuzumab is administered forsix 28-day cycles as follows: 100 mg on day 1, cycle 1; 900 mg on day 2cycle 1; 1000 mg on days 8 and 15 of cycle 1; and 1000 mg on day 1 ofcycles 2-6. Accordingly, in some embodiments, the term “rituximab”encompasses all corresponding anti-CD20 antibodies that fulfill therequirements necessary for obtaining a marketing authorization as anidentical or biosimilar product in a country or territory selected fromthe group of countries consisting of the USA, Europe and Japan.

In some embodiments, an anti-CD20 antibody has the same or similaractivity as rituximab, or a biosimilar thereof. In some embodiments, ananti-CD20 antibody binds to the same or similar region or epitope asrituximab or a fragment thereof. In some embodiments, an anti-CD20antibody competes with the binding of rituximab or a fragment thereof toCD20. In some embodiments, an anti-CD20 antibody is bioequivalent torituximab or a fragment thereof. In some embodiments, an anti-CD20antibody is a biosimilar of rituximab or a fragment thereof. In someembodiments, an anti-CD20 antibody is a variant or derivative ofrituximab, including functional fragments, derivatives, or antibodyconjugates.

Rituximab

Rituximab (Rituxan® or MabThera®) is a genetically engineered cytolytic,chimeric murine/human monoclonal IgG_(i) kappa antibody directed againstthe CD20 cell-surface molecule present in normal B lymphocytes andB-cell CLL and in most forms of non-Hodgkin's B-cell lymphomas.Rituximab has a binding affinity for the CD20 antigen of approximately8.0 nM. Rituximab can induce complement-dependent cellular cytotoxicity(CDC) and anti-body-dependent cellular cytotoxicity (ADCC), leading toits clinical activity against lymphoma cells. Rituximab can also lead toapoptosis of B cells upon binding to CD20, thereby leading to directinhibition of cellular growth.

Rituximab is produced by mammalian cell (Chinese Hamster Ovary)suspension culture in a nutrient medium containing the antibioticgentamicin. Gentamicin is not detectable in the final product. Rituximabis a sterile, clear, colorless, preservative-free liquid concentrate forintravenous administration. Rituximab is supplied at a concentration of10 mg/mL in either 100 mg/10 mL or 500 mg/50 mL single-use vials.Rituximab is formulated in polysorbate 80 (0.7 mg/mL), sodium citratedihydrate (7.35 mg/mL), sodium chloride (9 mg/mL) and water forinjection. The pH of Rituxan® (or MabThera®) is 6.5

Rituximab has been investigated in clinical studies and approved fortreatment of patients with CLL in combination with fludarabine andcyclophosphamide, as well as patients with rheumatoid arthritis incombination with methotrexate. Rituximab is also approved for treatmentof non-Hodgkin's lymphoma, Wegener's Granulomatosis and MicroscopicPolyangiitis.

In some embodiments, provided methods comprise administering to apatient in need thereof a combination of Compound 1 and rituximab,wherein the patient is further treated with fludarabine andcyclophosphamide in accordance with the approved indications.

I. General Dosing Protocol

As described herein, provided methods comprise administering Compound 1and an anti-CD20 antibody (e.g., rituximab, ofatumumab, obinutuzumab,etc.) to a patient in need thereof. Such methods optionally furthercomprise administering either (i) fludarabine and cyclophosphamide or(ii) bendamustine. It will be appreciated that each of the therapeuticagents (i.e., Compound 1, anti-CD20 antibody, fludarabine,cyclophosphamide and bendamustine) can be administered simultaneously orsequentially (e.g., Compound 1 can be administered before, during orafter an anti-CD20 antibody and/or either (i) fludarabine andcyclophosphamide or (ii) bendamustine and vice versa) as part of adosing regimen. For example, Compound 1 may be administered one or morehours, days or weeks before administration of an anti-CD20 antibody. Insome embodiments, Compound 1 and an anti-CD20 antibody may beadministered one or more hours, days or weeks before administration ofeither (i) fludarabine and cyclophosphamide or (ii) bendamustine.

In some embodiments, the present invention provides methods fortreating, stabilizing or lessening the severity or progression of one ormore diseases or conditions associated with BTK. In some embodiments,the present invention provides methods for preventing the progression ofa disease or disorder associated with BTK. In some embodiments, thedisease or disorder associated with BTK is selected from chroniclymphocytic leukemia (CLL) and small lymphocytic lymphoma (SLL).

In some embodiments, the disease or disorder associated with BTK isrefractory CLL. In some embodiments, the disease or disorder associatedwith BTK is relapsed CLL. In some embodiments, the disease or disorderassociated with BTK is refractory SLL. In some embodiments, the diseaseor disorder associated with BTK is relapsed SLL.

In some embodiments, provided methods comprise administering to apatient in need thereof Compound 1 in combination with rituximab. Insome such embodiments, each of Compound 1 and rituximab is administeredas a composition further comprising one or more pharmaceuticallyacceptable excipients. In some embodiments, provided methods compriseadministering each of Compound 1, rituximab, fludarabine andcyclophosphamide. In some such embodiments, each of Compound 1,rituximab, fludarabine and cyclophosphamide is administered as acomposition further comprising one or more pharmaceutically acceptableexcipients. In some embodiments, provided methods comprise administeringeach of Compound 1, rituximab and bendamustine. In some suchembodiments, each of Compound 1, rituximab and bendamustine isadministered as a composition further comprising one or morepharmaceutically acceptable excipients.

In some embodiments, provided methods comprise administering to apatient in need thereof a therapeutically effective amount of Compound 1in combination with a therapeutically effective amount of rituximab.Accordingly, in some embodiments, the present invention provides amethod of treating, stabilizing or lessening the severity or progressionof one or more diseases associated with BTK, the method comprisingadministering to a patient in need thereof a therapeutically effectiveamount of Compound 1 in combination with a therapeutically effectiveamount of rituximab. In some embodiments, provided methods compriseadministering to a patient in need thereof therapeutically effectiveamounts of each of Compound 1, rituximab, fludarabine andcyclophosphamide. In some embodiments, provided methods compriseadministering to a patient in need thereof therapeutically effectiveamounts of each of Compound 1, rituximab and bendamustine.

In some embodiments, provided methods comprise administering Compound 1in combination with rituximab, wherein Compound 1 is administered oncedaily (“QD”). In some embodiments, provided methods compriseadministering Compound 1 in combination with rituximab, wherein Compound1 is administered twice daily (“BID”). For purposes of clarity,administration of a 375 mg dose of Compound 1 “BID” means that thepatient is administered two separate doses of 375 mg in one day. In someembodiments, provided methods comprise administering each of Compound 1,rituximab, fludarabine and cyclophosphamide, wherein Compound 1 isadministered twice daily (“BID”). In some embodiments, provided methodscomprise administering each of Compound 1, rituximab and bendamustine,wherein Compound 1 is administered twice daily (“BID”).

In some embodiments, provided methods comprise administering Compound 1in combination with rituximab, wherein rituximab is administered onceduring a 28-day cycle. In some embodiments, provided methods compriseadministering Compound 1 in combination with rituximab, whereinrituximab is administered on cycle 1 day 1 or day 2. In someembodiments, provided methods comprise administering Compound 1 incombination with rituximab, wherein rituximab is administered on day 1of a 28-day cycle. In some such embodiments, rituximab is administeredon day 1 of cycles 2-6. In some embodiments, rituximab is administeredon day 1 of cycles 2-5. In some embodiments, rituximab is administeredon day 1 of cycles 2-4. In some embodiments, rituximab is administeredon day 1 of cycles 2-3. In some embodiments, provided methods compriseadministering each of Compound 1, rituximab, fludarabine andcyclophosphamide, wherein rituximab is administered once during a 28-daycycle. In some such embodiments, rituximab is administered on day 1 orday 2 of a 28-day cycle.

In some embodiments, provided methods comprise administering each ofCompound 1, rituximab and bendamustine, wherein rituximab isadministered once during a 28-day cycle. In some such embodiments,rituximab is administered on day 1 or day 2 of a 28-day cycle.

In some embodiments, provided methods comprise administering Compound 1in combination with rituximab, wherein Compound 1 is administered twicedaily and rituximab is administered once during a 28-day cycle. In someembodiments, provided methods comprise administering Compound 1 incombination with rituximab, wherein Compound 1 is administered twicedaily and rituximab is administered on cycle 1 day 1 or day 2. In someembodiments, provided methods comprise administering Compound 1 incombination with rituximab, wherein Compound 1 is administered twicedaily and rituximab is administered on day 1 of a 28-day cycle. In somesuch embodiments, rituximab is administered on day 1 of cycles 2-6. Insome embodiments, provided methods comprise administering each ofCompound 1, rituximab, fludarabine and cyclophosphamide, whereinCompound 1 is administered twice daily and rituximab is administeredonce during a 28-day cycle. In some such embodiments, rituximab isadministered on day 1 or day 2 of a 28-day cycle.

In some embodiments, provided methods comprise administering each ofCompound 1, rituximab and bendamustine, wherein Compound 1 isadministered twice daily and rituximab is administered once during a28-day cycle. In some such embodiments, rituximab is administered on day1 or day 2 of a 28-day cycle.

In some embodiments, each of Compound 1 and rituximab is administered aspharmaceutically acceptable compositions. In some embodiments, apharmaceutically acceptable composition comprising Compound 1 isformulated as an oral dosage form. In some embodiments, such oral dosageforms are capsules. In some embodiments, the pharmaceutically acceptablecomposition comprising rituximab is formulated as an intravenouscomposition. In some embodiments, fludarabine, cyclophosphamide andbendamustine are formulated as intravenous compositions.

In some embodiments, a pharmaceutically acceptable compositioncomprising Compound 1 comprises from about 5% to about 60% of Compound1, or a pharmaceutically acceptable salt thereof, based upon totalweight of the composition. In some embodiments, a pharmaceuticallyacceptable composition comprising Compound 1 comprises from about 5% toabout 15% or about 7% to about 15% or about 7% to about 10% or about 9%to about 12% of Compound 1, based upon total weight of the composition.In some embodiments, provided methods comprise administering to apatient in need thereof a pharmaceutically acceptable compositioncomprising from about 25% to about 75% or about 30% to about 60% orabout 40% to about 50% or about 40% to about 45% of Compound 1, basedupon total weight of the formulation. In certain embodiments, providedregimens comprise administering to a patient in need thereof apharmaceutically acceptable composition comprising from about 6%, about7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%,about 20%, about 30%, about 40%, about 41%, about 42%, about 43%, about44%, about 45%, about 50%, about 60%, about 70%, or about 75% ofCompound 1, based upon total weight of given composition or formulation.

Rituximab is commercially available as a 10 mg/mL solution comprisingsodium citrate, polysorbate 80, sodium chloride, sodium hydroxide,hydrochloric acid and water. Commercially available vials compriseeither 100 mg/10 mL or 500 mg/50 mL.

In some embodiments, a pharmaceutically acceptable composition comprisesfrom about 1 mg/mL to about 4 mg/mL rituximab. In some embodiments, apharmaceutically acceptable composition comprises from about 1 mg/mL,about 2 mg/mL, about 3 mg/mL or about 4 mg/mL rituximab. In someembodiments, a pharmaceutically acceptable composition comprises 10mg/mL.

Fludarabine (Fludara®) is commercially available as a vial of sterilelyophilized solid cake containing 50 mg of fludarabine phosphate, 50 mgof mannitol and sodium hydroxide to adjust pH to 7.7. The pH range forthe final solution is 7.2-8.2. The solid cake is reconstituted with 2 mLof Sterile Water for Injection USP, which results in a solutioncontaining 25 mg/mL of fludarabine phosphate intended for intravenousadministration.

Cyclophosphamide (Cytoxan®) is commercially available as a sterilepowder which may be prepared for parenteral use by infusion byreconstituting, for example, in 0.9% sterile sodium chloride (5 mL per100 mg anhydrous powder). Alternative solutions for reconstitution maybe found, for example, in the package insert.

Bendamustine (Treanda®) is commercially available as a single-use vialcontaining 100 mg of bendamustine hydrochloride as a lyophilized powder.The powder is reconstituted with 20 mL of Sterile Water for InjectionUSP to a final concentration of 5 mg/mL. Immediatly prior to infusion,the 5 mg/mL reconstituted solution is transferred to a 500 mL infusionbag containing 0.9% Sodium Chloride Injection USP. Alternatively, the 5mg/mL reconstituted solution may be transferred to a 500 mL infusion bagcontaining 2.5% Dextrose/0.45% Sodium Chloride Injection USP. The finalconcentration of bendamustine hydrochloride in the infusion bag shouldbe about 0.2-0.6 mg/mL.

In some embodiments, provided methods comprise administering Compound 1in combination with rituximab daily for a period of 1, 2, 3, 4, 5, 6, 7,8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27 or 28 days. In some embodiments, a treatment regimen comprises atleast one 28-day cycle. As used herein, the term “28-day cycle” meansthat provided treatment regimens are administered to a patient in needthereof for 28 consecutive days. In some embodiments, the combination ofCompound 1 and rituximab is administered for at least two, at leastthree, at least four, at least five or at least six 28-day cycles. Insome embodiments, the combination of Compound 1 and rituximab isadministered for at least seven, at least eight, at least nine, at leastten, at least eleven or at least twelve 28-day cycles. In someembodiments, the combination of Compound 1 and rituximab is administeredfor at least thirteen, at least fourteen, at least fifteen, at leastsixteen, at least seventeen, at least eighteen, at least nineteen or atleast twenty 28-day cycles. In some embodiments, the combination ofCompound 1 and rituximab is administered to a patient for the durationof the patient's life.

In some embodiments, provided methods comprise administering to apatient in need thereof each of Compound 1, rituximab, fludarabine andcyclophosphamide, wherein each of rituximab, fludarabine andcyclophosphamide is administered for at least one, two, at least three,at least four, at least five or at least six 28-day cycles.

In some embodiments, the combination of Compound 1 and rituximab isadministered for at least six 28-day cycles, and Compound 1 isadministered for at least one additional 28-day cycle. In someembodiments, the combination of Compound 1 and rituximab is administeredfor at least six 28-day cycles, and Compound 1 is administered for anadditional two, three, four, five, six, seven, eight, nine, ten, eleven,twelve, thirteen or fourteen 28-day cycles. In some embodiments, thecombination of Compound 1 and rituximab is administered for at least six28-day cycles, and Compound 1 is administered for the duration of thepatient's life. In some embodiments, Compound 1 is administered on days1 through 28 (for example, one dose each day or two doses each day) ofone or more 28-day cycles and rituximab is administered on day 1 of a28-day cycle. In some embodiments, Compound 1 is administered on days 1through 28 of one or more 28-day cycles and rituximab is administered onday 1 or day 2 of a 28-day cycle.

In some embodiments, two adjacent 28-day cycles may be separated by arest period. Such a rest period may be one, two, three, four, five, six,seven or more days during which the patient is not administered eitheror both Compound 1 and rituximab. In a preferred embodiment, twoadjacent 28-day cycles are continuous.

In some embodiments, provided methods comprise administering to apatient in need thereof Compound 1 in combination with rituximab,wherein the patient has failed at least one prior therapy. In someembodiments, provided methods comprise administering to a patient inneed thereof Compound 1, rituximab, fludarabine and cyclophosphamide,wherein the patient has failed at least one prior therapy. In someembodiments, provided methods comprise administering to a patient inneed thereof Compound 1, rituximab and benmustine, wherein the patienthas failed at least one prior therapy.

Unit Dosage Forms

Pharmaceutical compositions for use in the present invention may beprepared as a unit dosage form. A person of ordinary skill willappreciate that the unit dosage forms described herein refer to anamount of a component in its free base form. A person skilled in the artwill further appreciate that, when a pharmaceutical compositioncomprises a salt form of one component, for example, a besylate saltform of Compound 1, the amount of the salt form present in thecomposition is an amount that is equivalent to a unit dose of the freebase of the component (i.e., of Compound 1). For example, apharmaceutical composition comprising a besylate salt of Compound 1would contain 34.97 mg of the besylate salt form necessary to deliver anequivalent 25 mg unit dose of the free base of Compound 1.

In some embodiments, provided methods comprise administering to apatient in need thereof a therapeutically effective amount of Compound1, wherein the therapeutically effective amount of Compound 1 is about250 mg to about 1250 mg. In some embodiments, the therapeuticallyeffective amount of Compound 1 is administered as one or more discreetdoses. For example, in some embodiments, a therapeutically effectiveamount of Compound 1 is 250 mg, wherein the therapeutically effectiveamount is administered as 125 mg twice daily (BID). In some embodiments,a therapeutically effective amount of Compound 1 is 500 mg, wherein thetherapeutically effective amount is administered as 250 mg twice daily(BID). In some embodiments, a therapeutically effective amount ofCompound 1 is 750 mg, wherein the therapeutically effective amount isadministered as 375 mg twice daily (BID). In some embodiments, atherapeutically effective amount of Compound 1 is 1000 mg, wherein thetherapeutically effective amount is administered as 500 mg twice daily(BID).

In some embodiments, provided methods comprise administering to apatient in need thereof a therapeutically effective amount of Compound1, wherein the therapeutically effective amount of Compound 1 is about125 mg to about 1250 mg, or about 125 mg to about 1125 mg, or about 125mg to about 1000 mg, or about 125 mg to about 875 mg, or about 125 mg toabout 750 mg, or about 125 mg to about 625 mg, or about 125 mg to about500 mg, or about 125 mg to about 375 mg, or about 125 mg to about 250mg, or about 250 mg to about 1250 mg, or about 250 mg to about 1125 mg,or about 250 mg to about 1000 mg, or about 250 mg to about 875 mg, orabout 250 mg to about 750 mg, or about 250 mg to about 625 mg, or about250 mg to about 500 mg, or about 250 mg to about 375 mg, or about 375 mgto about 1250 mg, or about 375 mg to about 1125 mg, or about 375 mg toabout 1000 mg, or about 375 mg to about 875 mg, or about 375 mg to about750 mg, or about 375 mg to about 625 mg, or about 375 mg to about 500mg, or about 500 mg to about 1250 mg, or about 500 mg to about 1125 mg,or about 500 mg to about 1000 mg, or about 500 mg to about 750 mg, orabout 500 mg to about 625 mg, or about 625 mg to about 1250 mg, or about625 mg to about 1125 mg, or about 625 mg to about 1000 mg, or about 625mg to about 875 mg, or about 625 mg to about 750 mg, or about 750 mg toabout 1250 mg, or about 750 mg to about 1125 mg, or about 750 mg toabout 1000 mg, or about 875 mg to about 1250 mg, or about 875 mg toabout 1125 mg, or about 875 mg to about 1000 mg.

In some embodiments, provided methods comprise administering to apatient in need thereof a therapeutically effective amount of Compound1, wherein the therapeutically effective amount of Compound 1 is about125 mg, 130 mg, 135 mg, 140 mg, 145 mg, 150 mg, 155 mg, 160 mg, 165 mg,170 mg, 175 mg, 180 mg, 185 mg, 190 mg, 195 mg, 200 mg, 205 mg, 210 mg,215 mg, 220 mg, 225 mg, 230 mg, 235 mg, 240 mg, 245 mg, 250 mg, 255 mg,260 mg, 265 mg, 270 mg, 275 mg, 280 mg, 285 mg, 290 mg, 295 mg, 300 mg,305 mg, 310 mg, 315 mg, 320 mg, 325 mg, 330 mg, 335 mg, 340 mg, 345 mg,350 mg, 355 mg, 360 mg, 365 mg, 370 mg, 375 mg, 380 mg, 385 mg, 390 mg,395 mg, 400 mg, 405 mg, 410 mg, 415 mg, 420 mg, 425 mg, 430 mg, 435 mg,440 mg, 445 mg, 450 mg, 455 mg, 460 mg, 465 mg, 470 mg, 475 mg, 480 mg,485 mg, 490 mg, 495 mg, 500 mg, 505 mg, 510 mg, 515 mg, 520 mg, 525 mg,530 mg, 535 mg, 540 mg, 545 mg, 550 mg, 555 mg, 560 mg, 565 mg, 570 mg,575 mg, 580 mg, 585 mg, 590 mg, 595 mg, 600 mg, 605 mg, 610 mg, 615 mg,620 mg, 625 mg, 630 mg, 635 mg, 640 mg, 645 mg, 650 mg, 655 mg, 660 mg,665 mg, 670 mg, 675 mg, 680 mg, 685 mg, 690 mg, 695 mg, 700 mg, 705 mg,710 mg, 715 mg, 720 mg, 725 mg, 730 mg, 735 mg, 740 mg, 745 mg, 750 mg,755 mg, 760 mg, 765 mg, 770 mg, 775 mg, 780 mg, 785 mg, 790 mg, 795 mg,800 mg, 805 mg, 810 mg, 815 mg, 820 mg, 825 mg, 830 mg, 835 mg, 840 mg,845 mg, 850 mg, 855 mg, 860 mg, 865 mg, 870 mg, 875 mg, 880 mg, 885 mg,890 mg, 895 mg, 900 mg, 905 mg, 910 mg, 915 mg, 920 mg, 925 mg, 930 mg,935 mg, 940 mg, 945 mg, 950 mg, 955 mg, 960 mg, 965 mg, 970 mg, 975 mg,980 mg, 985 mg, 990 mg, 995 mg, 1000 mg, 1005 mg, 1010 mg, 1015 mg, 1020mg, 1025 mg, 1030 mg, 1035 mg, 1040 mg, 1045 mg, 1050 mg, 1055 mg, 1060mg, 1065 mg, 1070 mg, 1075 mg, 1080 mg, 1085 mg, 1090 mg, 1095 mg, 1100mg, 1105 mg, 1110 mg, 1115 mg, 1120 mg, 1125 mg, 1130 mg, 1135 mg, 1140mg, 1145 mg, 1150 mg, 1155 mg, 1160 mg, 1165 mg, 1170 mg, 1175 mg, 1180mg, 1185 mg, 1190 mg, 1195 mg, 1200 mg, 1205 mg, 1210 mg, 1215 mg, 1220mg, 1225 mg, 1230 mg, 1235 mg, 1240 mg, 1245 mg or 1250 mg.

In some embodiments, provided methods comprise administering to apatient in need thereof a pharmaceutical composition comprising a unitdose of Compound 1 in combination with rituximab. In some suchembodiments, the unit dose of Compound 1 is about 25 mg, about 50 mg,about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg,about 200 mg, about 225 mg or about 250 mg.

In some embodiments, provided methods comprise administering to apatient in need thereof a pharmaceutical composition comprisingrituximab, wherein rituximab is administered as an infusion at a rate of50 mg/hr. In some embodiments, the infusion rate of rituximab isincreased by 50 mg/hr every 30 minutes, to a maximum of 400 mg/hr. Insome embodiments, the infusion rate of rituximab is increased by 100mg/hr every 30 minutes, to a maximum of 400 mg/hr. Accordingly, in someembodiments, the infusion rate of rituximab is 100 mg/hr. In someembodiments, the infusion rate of rituximab is 150 mg/hr. In someembodiments, the infusion rate of rituximab is 200 mg/hr. In someembodiments, the infusion rate of rituximab is 250 mg/hr. In someembodiments, the infusion rate of rituximab is 300 mg/hr. In someembodiments, the infusion rate of rituximab is 350 mg/hr. In someembodiments, the infusion rate of rituximab is 400 mg/hr.

II. Uses of Compounds and Pharmaceutically Acceptable Compositions

Compound 1 and compositions described herein are generally useful forthe inhibition of protein kinase activity of one or more enzymes.Examples of kinases that are inhibited by Compound 1 and compositionsdescribed herein and against which the methods described herein areuseful include BTK and other TEC-kinases, including ITK, TEC, BMX andRLK, or a mutant thereof.

Bruton's tyrosine kinase (“BTK”), a member of TEC-kinases, is a keysignaling enzyme expressed in B-lymphocytes, monocytes, and mast cellsor basophils. BTK plays an essential role in the B-cell signalingpathway linking cell surface B-cell receptor (BCR) stimulation todownstream intracellular responses.

BTK is a key regulator of B-cell development, activation, signaling, andsurvival (Kurosaki, Curr. Op. Imm., 2000, 276-281; Schaeffer andSchwartzberg, Curr. Op. Imm. 2000, 282-288). In addition, BTK plays arole in a number of other hematopoietic cell signaling pathways, e.g.,Toll like receptor (TLR) and cytokine receptor-mediated TNF-α productionin macrophages, IgE receptor (Fc_epsilon_RI) signaling in mast cells,inhibition of Fas/APO-1 apoptotic signaling in B-lineage lymphoid cells,and collagen-stimulated platelet aggregation. See, e.g., C. A. Jeffries,et al., (2003), Journal of Biological Chemistry 278:26258-26264; N. J.Horwood, et al., (2003), The Journal of Experimental Medicine 197:1603-1611; Iwaki et al. (2005), Journal of Biological Chemistry280(48):40261-40270; Vassilev et al. (1999), Journal of BiologicalChemistry 274(3): 1646-1656, and Quek et al. (1998), Current Biology8(20): 1137-1140.

Patients with inherited inactivating mutations in BTK have a profoundblock in B-cell development, resulting in the almost complete absence ofmature B lymphocytes and plasma cells, severely reduced Ig levels and aprofound inhibition of humoral response to recall antigens (reviewed inVihinen et al Frontiers in Bioscience 5: d917-928). Mice deficient inBTK also have a reduced number of peripheral B-cells and greatlydecreased serum levels of IgM and IgG3. BTK deletion in mice has aprofound effect on B-cell proliferation induced by anti-IgM, andinhibits immune responses to thymus-independent type II antigens(Ellmeier et al, J Exp Med 192: 1611-1623 (2000)). BTK also plays acrucial role in mast cell activation through the high-affinity IgEreceptor (Fc_epsilon_RI). BTK deficient murine mast cells have reduceddegranulation and decreased production of proinflammatory cytokinesfollowing Fc_epsilon_RI cross-linking (Kawakami et al. Journal ofLeukocyte Biology 65: 286-290).

Compound 1 is an inhibitor of BTK and therefore useful for treating oneor more disorders associated with activity of BTK. Thus, in someembodiments, the present invention provides a method of treating,stabilizing or lessening the severity or progression of a BTK-mediateddisorder comprising the step of administering to a patient in needthereof Compound 1 in combination with rituximab.

Chronic Lymphocytic Leukemia and Small Lymphocytic Lymphoma

The B-cell disorders chronic lymphocytic leukemia (CLL) and smalllymphocytic lymphoma (SLL) represent two ends of a spectrum of the samedisease process differing in the degree of blood/marrow involvement(CLL) versus lymph node involvement (SLL). CLL is a lymphoproliferativemalignancy characterized by progressive accumulation of morphologicallymature but functionally incompetent lymphocytes in the blood, bonemarrow, and lymphoid tissues. It affects mainly elderly individuals withthe median age at presentation of 65 to 70 years. The clinical course ofCLL ranges from indolent disease with long-term survival over 12 yearsto aggressive disease with median survival of 2 years.

Chronic lymphocytic leukemia is the most common leukemia in the U.S. andis typically characterized immunophenotypically as CD5+, CD23+, CD10−,CD19+, CD20 dim, sIg dim, and cyclin D1- (the latter point adistinguishing feature from mantle cell lymphoma). Chronic lymphocyticleukemia must also be distinguished from monoclonal B lymphocytosis(absolute monoclonal B-cell count <5000/μL and absence of adenopathy orother clinical features of lymphoproliferative disorder). Theunderstanding of CLL/SLL biology and prognostic factors, and advances informulating a risk-stratified approach to treatment of CLL/SLL have beenrecently reviewed by Lanasa, Furman, and the National ComprehensiveCancer Network NHL panel.

The cellular expression of Btk is restricted and largely limited toB-lymphocytes, monocytes, and mast cells or basophils. Investigation hasrevealed that some B-cell lymphomas and CLL/SLL depend on BCR signaling,suggesting that interruption of such signaling could be a promisingtherapeutic opportunity Recently it has been reported that half of allCLL retain BCR signaling in vitro and that immunoglobulin heavy genesomatic mutation (IgVH) is an important determinant of BCRresponsiveness. Indeed, the mutational status of the BCR in CLL is oneof the strongest predictors of disease progression, as aggressivedisease typically displays BCR encoded by unmutated immunoglobulinvariable heavy chains.

Two groups have reported that mutated and unmutated CLL cells responddifferentially to IgM ligation of the BCR, with unmutated, but notmutated, CLL cells responding to BCR stimulation with increased globaltyrosine phosphorylation and by up-regulating several genes associatedwith cell cycle regulation and allowing cell growth and expansion. Thesedata highlight the differential role that BCR signaling plays in CLLphysiology depending on IgVH mutational status and may suggest apossible differential responsiveness of CLL to inhibitors of BCRsignaling. Other in vitro studies have reported that specific Btkinhibition with the investigational agent PCI-32765 producedsubstantially more apoptosis and cytotoxicity in CLL cells relative tonormal B-cells; as well as inducing apoptosis in the face ofanti-apoptotic micro-environmental signals, reduction of secretion ofchemokines CCL3 and CCL4, and reduction of chemotaxis towards thechemokines CXCL12 and CXCL13. Detailed studies of the pathophysiologicrole of Btk in the origin and/or maintenance of Waldenstrom'smacroglobulinemia (WM) have not yet been reported. However, a recentreport investigating transgenic mouse models demonstrated thatconstitutively active Btk expression resulted in selective expansion orsurvival of B-1 cells that were driven into germinal center independentplasma cell differentiation, as evidenced by increased numbers of IgM+plasma cells in spleen and bone marrow and significantly elevated serumIgM. Anti-nucleosome autoantibodies and glomerular IgM deposition werealso observed. However, one study of sequence analysis in 19 WM patientswith hypogammaglobulinemia G and/or A failed to find any novel variantsin the promoter, flanking introns, or exons of Btk.

Allogeneic stem cell transplant is the only potentially curativetreatment for CLL, but 70% of affected patients are ≧65 years of age atthe time of diagnosis, have co-morbid conditions limiting eligibilityfor such therapy, and may exhibit a prolonged natural history with orwithout specific treatment. The actual prognosis of CLL is variable anddependent principally on clinical stage and certain genetic andmolecular features. Both the Rai and Binet clinical staging systems areable to distinguish patient prognostic groups with median OSs rangingfrom 19 months in the most advanced stage (thrombocytopenia) to >150months in the earliest stage (blood and marrow lymphocytosis withoutadenopathy, organomegaly, or defined anemia/thrombocytopenia).Classification by the presence or absence of IgVH and by interphasefluorescent in situ hybridization (iFISH) analysis for probed-foracquired chromosomal abnormalities adds additional prognosticdiscrimination to clinical staging, with unmutated IgVH and del(11q) anddel(17p) cytogenetics predicting poorer outcome.

The CLL treatment algorithm is complex and requires first the decisionto treat (e.g., presence of symptoms such as fatigue or night sweats;bulky adenopathy/organomegaly; progressive anemia/thrombocytopenia); andsecond, choice of the treatment regimen, usually involving one or more:purine nucleosides (fludarabine), alkylating agents (cyclophosphamide,chlorambucil, bendamustine), corticosteroids, anti-CD20 monoclonalantibodies (rituximab/ofatumumab), or anti-CD52 monoclonal antibodies(alemtuzumab). The choice of specific therapies depends on the patient'sage, disease pattern (eg, primarily nodal versus non-nodal), anticipateddrug tolerance and contraindications, and presence or absence of adverseprognostic features such as del(11q) or del(17p). Despite numeroustherapies, treatment options are eventually limited by drug toxicitiesand resistance, and patients who do not succumb to other maladies endureprogressive complications relating to cytopenias, the effects oflymphadenopathy and organomegaly, systemic symptoms, and infectiouscomplications. Given the often elderly character of the patientpopulation, an orally available, well tolerated treatment that exploitsa novel weakness of CLL would be welcome.

Rationale for Targeting Btk and Combinations with Rituximab in CLL andSLL

Strategies specifically targeting B-cells, for example the B-celldepleting anti-CD20 monoclonal antibodies rituximab and ofatumumab, havedemonstrated clinical efficacy in B-cell lymphoma and CLL. Spleentyrosine kinase (Syk) is a kinase in the BCR signaling pathway proximalto Btk. Inhibition of Syk with the orally available Syk inhibitorfostamatinib disodium produced clinical responses in DLBCL, CLL, andmantle cell lymphoma. Most tellingly, clinical proof of concept for Btkinhibition has been demonstrated by clinical investigations of theorally available Btk inhibitor PCI-32765, which have reported objectiveanti-tumor responses in patients with DLBCL; mantle cell, marginalzone/mucosa-associated lymphoid tissue (MALT), and follicular lymphoma(FL), WM, and CLL/SLL, with good tolerability.

Thus, based on the critical importance of BCR signaling mediated throughBtk for the survival and proliferation of various malignant B-cells;Btk's limited cellular expression in B-cells, macrophages, andmonocytes; and demonstrated pre-clinical and early clinical proofs ofconcept that Btk inhibition produces salutary anti-lymphoma, CLL, and WMeffects with acceptable clinical tolerability, targeting Btk with aselective Btk inhibitor is a promising and appropriate therapeuticstrategy to investigate further in the clinic. Compound 1, as itsbesylate salt, has been shown in recent studies to be safe and effectiveagainst CLL as a single agent therapeutic. As of Sep. 11, 2012, 35 outof 43 patients with CLL and have experienced stable disease and continueto on treatment withN-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamidebesylate monotherapy. 15 of 23 patients have experienced reductions inlymph node size, and 28 of 33 patients have experienced early increasesin absolute lymphocyte counts (ALC). See U.S. patent application Ser.No. 13/661,678 and International Patent Application No.PCT/US2012/062133, both filed on Oct. 26, 2012, each of which is herebyincorporated by reference in its entirety. Such data strongly supportthe use of a BTK inhibitor, and Compound 1 in particular, for treatingCLL. Compound 1 is generally well tolerated as a single agent at up to750 mg PO QD and the maximum tolerated dose (MTD) has not yet beenreached. Studies are ongoing and additional dose levels currently beinginvestigated include: 1000 mg QD, 1250 mg QD, 375 mg BID and 500 mg BID.

Rituximab has also been shown to exhibit good activity againstrelapsed/refractory CLL patients. In one study, rituximab, incombination with fludarabine/cyclophosphamide, was evaluated in 408patients with CLL and showed an 86% response rate, as compared to the73% response rate observed for fludarabine/cyclophosphamide alone. Themedian progression-free survival was 39.8 months, as compared to 31.5months observed for fludarabine/cyclophosphamide alone. Accordingly, insome embodiments, the present invention encompasses the recognition thata BTK inhibitor such as Compound 1 in combination with rituximab isuseful in the treatment of CLL and SLL. Compound 1 either as a singleagent or in combination, may be found to be efficacious in CLL patients,including but not limited to those who had expressed one or more of thefollowing prognostic/genetic markers and cytogenetic risk factors:deletions of chromosome 11q, 17p or 13q, or Trisomy 12 and 14q,zeta-chain-associated protein kinase 70 (ZAP 70) or immunoglobulin heavychain variable region (IgVH) un-mutated.

In some embodiments, the present invention provides methods of treating,stabilizing or lessening the severity or progression of one or morediseases and conditions associated with BTK comprising administering toa patient in need thereof Compound 1 in combination with rituximab.

III. Methods of Treating Diseases or Disorders Associated with Btk

In some embodiments, the present invention provides a method oftreating, stabilizing or lessening the severity or progression of adisease or disorder selected from the group consisting of chroniclymphocytic leukemia and small lymphocytic lymphoma, the methodcomprising administering to a patient in need thereof Compound 1 incombination with rituximab. In some embodiments, provided methodscomprise administering to a patient in need thereof each of Compound 1,rituximab, fludarabine and cyclophosphamide. In some embodiments,provided methods comprise administering to a patient in need thereofeach of Compound 1, rituximab and bendamustine.

In some embodiments, provided methods comprise administering to apatient in need thereof a composition comprising Compound 1 incombination with a composition comprising rituximab. In someembodiments, the composition comprising Compound 1 further comprises oneor more pharmaceutically acceptable excipients. In some suchembodiments, the composition comprising Compound 1 is formulated as anoral dosage form. In some embodiments, the oral dosage form is acapsule. In some embodiments, provided methods comprise administering toa patient in need thereof compositions comprising each of Compound 1,rituximab, fludarabine and cyclophosphamide. In some embodiments,provided methods comprise administering to a patient in need thereofcompositions comprising each of Compound 1, rituximab and bendamustine.

In some embodiments, the composition comprising rituximab furthercomprises one or more pharmaceutically acceptable excipients. In somesuch embodiments, the composition comprising rituximab is formulated asan intravenous dosage form.

In some embodiments, provided methods comprise administering to apatient in need thereof a unit dose of Compound 1 in combination with aunit dose of rituximab. In some embodiments, the unit dose of Compound 1is about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 125 mg,about 150 mg, about 175 mg, about 200 mg, about 225 mg or about 250 mg.

In some embodiments, provided methods comprise administering to apatient in need thereof a pharmaceutical composition comprisingrituximab, wherein rituximab is administered as an infusion at a rate of50 mg/hr. In some embodiments, the infusion rate of rituximab isincreased by 50 mg/hr every 30 minutes, to a maximum of 400 mg/hr. Insome embodiments, the infusion rate of rituximab is increased by 100mg/hr every 30 minutes, to a maximum of 400 mg/hr. Accordingly, in someembodiments, the infusion rate of rituximab is 100 mg/hr. In someembodiments, the infusion rate of rituximab is 150 mg/hr. In someembodiments, the infusion rate of rituximab is 200 mg/hr. In someembodiments, the infusion rate of rituximab is 250 mg/hr. In someembodiments, the infusion rate of rituximab is 300 mg/hr. In someembodiments, the infusion rate of rituximab is 350 mg/hr. In someembodiments, the infusion rate of rituximab is 400 mg/hr.

In some embodiments, the present invention provides a method oftreating, stabilizing or lessening the severity or progression of adisease or disorder selected from the group consisting of chroniclymphocytic leukemia and small lymphocytic lymphoma, the methodcomprising administering to a patient in need thereof Compound 1 incombination with rituximab, wherein the patient has failed at least oneprior therapy. In some embodiments, provided methods compriseadministering to a patient in need thereof Compound 1, rituximab,fludarabine and bendamustine, wherein the patient has failed at leastone prior therapy. In some embodiments, provided methods compriseadministering to a patient in need thereof Compound 1, rituximab andbendamustine, wherein the patient has failed at least one prior therapy.

In some embodiments, provided methods comprise administering to apatient in need thereof about 500 mg to about 1250 mg Compound 1 incombination with about 375 mg/m² to about 500 mg/m² rituximab. In someembodiments, provided methods comprise administering to a patient inneed thereof about 750 mg to about 1000 mg Compound 1 and about 375mg/m² to about 500 mg/m² rituximab. In some embodiments, providedmethods comprise administering to a patient in need thereof about 500 mgto about 1250 mg Compound 1, about 375 mg/m² to about 500 mg/m²rituximab, about 25 mg/m² fludarabine and about 250 mg/m²cyclophosphamide. In some embodiments, provided methods compriseadministering to a patient in need thereof about 750 mg to about 1000 mgCompound 1, about 375 mg/m² to about 500 mg/m² rituximab and about 70mg/m² bendamustine.

In some embodiments, the present invention provides a method oftreating, stabilizing or lessening the severity or progression of adisease or disorder selected from the group consisting of chroniclymphocytic leukemia and small lymphocytic lymphoma, the methodcomprising administering to a patient in need thereof about 375 mg BIDto about 500 mg BID Compound 1 in combination with about 375 mg/m² toabout 500 mg/m² rituximab. In some such embodiments, rituximab isadministered once during a 28-day cycle. In some embodiments, providedmethods comprise administering to a patient in need thereof about 250 mgto about 500 mg BID Compound 1, about 375 mg/m² to about 500 mg/m²rituximab, about 25 mg/m² fludarabine and about 250 mg/m²cyclophosphamide, wherein rituximab is administered on day 1 of a 28-daycycle. In some such embodiments, each of fludarabine andcyclophosphamide is administered on days 1-3 of a 28-day cycle. In someembodiments, provided methods comprise administering to a patient inneed thereof about 750 mg to about 1000 mg Compound 1, about 375 mg/m²to about 500 mg/m² rituximab and about 70 mg/m² bendamustine, whereinrituximab is administered on day 1 of a 28-day cycle. In some suchembodiments, bendamustine is administered on days 1 and 2 of a 28-daycycle.

In some embodiments, provided methods comprise administering to apatient in need thereof about 125 mg BID Compound 1 and about 375 mg/m²rituximab. In some such embodiments, provided methods further compriseadministering about 25 mg/m² fludarabine and about 250 mg/m²cyclophosphamide. In some embodiments, provided methods compriseadministering to a patient in need thereof about 125 mg BID Compound 1,about 375 mg/m² rituximab and about 70 mg/m² bendamustine.

In some embodiments, provided methods comprise administering to apatient in need thereof about 125 mg BID Compound 1 and about 500 mg/m²rituximab. In some such embodiments, provided methods further compriseadministering about 25 mg/m² fludarabine and about 250 mg/m²cyclophosphamide. In some embodiments, provided methods compriseadministering to a patient in need thereof about 125 mg BID Compound 1,about 500 mg/m² rituximab and about 70 mg/m² bendamustine.

In some embodiments, provided methods comprise administering to apatient in need thereof about 250 mg BID Compound 1 and about 375 mg/m²rituximab. In some such embodiments, provided methods further compriseadministering about 25 mg/m² fludarabine and about 250 mg/m²cyclophosphamide. In some embodiments, provided methods compriseadministering to a patient in need thereof about 250 mg BID Compound 1,about 375 mg/m² rituximab and about 70 mg/m² bendamustine.

In some embodiments, provided methods comprise administering to apatient in need thereof about 250 mg BID Compound 1 and about 500 mg/m²rituximab. In some such embodiments, provided methods further compriseadministering about 25 mg/m² fludarabine and about 250 mg/m²cyclophosphamide. In some embodiments, provided methods compriseadministering to a patient in need thereof about 250 mg BID Compound 1,about 500 mg/m² rituximab and about 70 mg/m² bendamustine.

In some embodiments, provided methods comprise administering to apatient in need thereof about 375 mg BID Compound 1 and about 375 mg/m²rituximab. In some such embodiments, provided methods further compriseadministering about 25 mg/m² fludarabine and about 250 mg/m²cyclophosphamide. In some embodiments, provided methods compriseadministering to a patient in need thereof about 375 mg BID Compound 1,about 375 mg/m² rituximab and about 70 mg/m² bendamustine.

In some embodiments, provided methods comprise administering to apatient in need thereof about 375 mg BID Compound 1 and about 500 mg/m²rituximab. In some such embodiments, provided methods further compriseadministering about 25 mg/m² fludarabine and about 250 mg/m²cyclophosphamide. In some embodiments, provided methods compriseadministering to a patient in need thereof about 375 mg BID Compound 1,about 500 mg/m² rituximab and about 70 mg/m² bendamustine.

In some embodiments, provided methods comprise administering to apatient in need thereof about 500 mg BID Compound 1 and about 375 mg/m²rituximab. In some such embodiments, provided methods further compriseadministering about 25 mg/m² fludarabine and about 250 mg/m²cyclophosphamide. In some embodiments, provided methods compriseadministering to a patient in need thereof about 500 mg BID Compound 1,about 375 mg/m² rituximab and about 70 mg/m² bendamustine.

In some embodiments, provided methods comprise administering to apatient in need thereof about 500 mg BID Compound 1 and about 500 mg/m²rituximab. In some such embodiments, provided methods further compriseadministering about 25 mg/m² fludarabine and about 250 mg/m²cyclophosphamide. In some embodiments, provided methods compriseadministering to a patient in need thereof about 500 mg BID Compound 1,about 500 mg/m² rituximab and about 70 mg/m² bendamustine.

In some embodiments, rituximab is administered once during a 28-daycycle. In some embodiments, rituximab is administered on cycle 1 day 1or day 2. In some embodiments, rituximab is administered on day 1 of a28-day cycle. In some embodiments, rituximab is administered on cycle 2day 1. In some embodiments, rituximab is administered on cycle 3 day 1.In some embodiments, rituximab is administered on cycle 4 day 1. In someembodiments, rituximab is administered on cycle 5 day 1. In someembodiments, rituximab is administered on cycle 6 day 1. In someembodiments, rituximab is administered on each of cycle 1 day 1 or day2, cycle 2 day 1, cycle 3 day 1, cycle 4 day 1, cycle 5 day 1 and cycle6 day 1.

In some embodiments, 375 mg/m² rituximab is administered on cycle 1 day1 or day 2, and 500 mg/m² rituximab is administered on cycle 2 day 1. Insome embodiments, 375 mg/m² rituximab is administered on cycle 1 day 1or day 2, and 500 mg/m² rituximab is administered on each of cycle 2 day1 and cycle 3 day 1. In some embodiments, 375 mg/m² rituximab isadministered on cycle 1 day 1 or day 2, and 500 mg/m² rituximab isadministered on each of cycle 2 day 1, cycle 3 day 1 and cycle 4 day 1.In some embodiments, 375 mg/m² rituximab is administered on cycle 1 day1 or day 2, and 500 mg/m² rituximab is administered on each of cycle 2day 1, cycle 3 day 1, cycle 4 day 1 and cycle 5 day 1. In someembodiments, 375 mg/m² rituximab is administered on cycle 1 day 1 or day2, and 500 mg/m² rituximab is administered on each of cycle 2 day 1,cycle 3 day 1, cycle 4 day 1, cycle 5 day 1 and cycle 6 day 1.

In some embodiments, 25 mg/m² fludarabine is administered on days 1-3 ofcycles 1, 2, 3, 4, 5 and/or 6. In some embodiments, 250 mg/m²cyclophosphamide is administered on days 1-3 1 of cycles 1, 2, 3, 4, 5and/or 6. In some embodiments, 70 mg/m² bendamustine is administered ondays 1 and 2 of cycles 1, 2, 3, 4, 5 and/or 6.

In some embodiments, the combination of Compound 1 and rituximab isadministered over a period of 28 consecutive days (“a 28-day cycle”). Insome embodiments, the combination of Compound 1 and rituximab isadministered for two, three, four, five or six 28-day cycles. In someembodiments, the combination of Compound 1 and rituximab is administeredfor one, two, three, four, five or six 28-day cycles, and Compound 1 isadministered for an additional one, two, three, four, five, six, seven,eight, nine, ten, eleven, twelve, thirteen or fourteen 28-day cycles. Insome embodiments, the combination of Compound 1 and rituximab isadministered to a patient for one, two, three, four, five or six 28-daycycles, and Compound 1 is administered for the duration of the patient'slife. In some embodiments, the combination of Compound 1 and rituximabis administered to a patient for one, two, three, four, five or six28-day cycles, and either of Compound 1 or rituximab is furtheradministered to the patient for one or more additional 28-day cycles. Insome embodiments, the combination of Compound 1 and rituximab isadministered to a patient for the duration of the patient's life.

In some embodiments, each of Compound 1, rituximab, fludarabine andcyclophosphamide is administered for one, two, three, four, five or six28-day cycles, and Compound 1 is administered for an additional one,two, three, four, five, six, seven, eight, nine, ten, eleven, twelve,thirteen or fourteen 28-day cycles.

In some embodiments, each of Compound 1, rituximab and bendamustine isadministered for one, two, three, four, five or six 28-day cycles, andCompound 1 is administered for an additional one, two, three, four,five, six, seven, eight, nine, ten, eleven, twelve, thirteen or fourteen28-day cycles.

In some embodiments, two adjacent 28-day cycles may be separated by arest period. Such a rest period may be one, two, three, four, five, six,seven or more days during which the patient is not administered eitheror both Compound 1 and rituximab. In a preferred embodiment, twoadjacent 28-day cycles are continuous.

In some embodiments, provided methods comprise administering to apatient in need thereof Compound 1 in combination with rituximab,wherein the patient has failed at least one prior therapy. In someembodiments, provided methods comprise administering to a patient inneed thereof each of Compound 1, rituximab, fludarabine andcyclophosphamide, wherein the patient has failed at least one priortherapy. In some embodiments, provided methods comprise administering toa patient in need thereof each of Compound 1, rituximab andbendamustine, wherein the patient has failed at least one prior therapy.

In some embodiments, the present invention provides a system fortreating, stabilizing or lessening the severity of one or more diseasesor conditions associated with BTK, the system comprising Compound 1 andrituximab. In some embodiments, the system is a kit. In some suchembodiments, the kit comprises a pharmaceutical composition comprisingCompound 1 and a pharmaceutical composition comprising rituximab.

In some embodiments, the kit comprises twenty-eight (28) daily doses ofCompound 1 and one 10 mg/mL vial of rituximab. In some embodiments, thekit comprises twenty-eight (28) daily doses of Compound 1 and one 100mg/10 mL vial of rituximab. In some embodiments, the kit comprisestwenty-eight (28) daily doses of Compound 1 and one 500 mg/50 mL vial ofrituximab.

In some embodiments, the kit comprises fifty-six (56) 375 mg doses ofCompound 1 and one 10 mg/mL vial of rituximab. In some embodiments, thekit comprises fifty-six (56) 375 mg doses of Compound 1 and one 100mg/10 mL vial of rituximab. In some embodiments, the kit comprisesfifty-six (56) 375 mg doses of Compound 1 and one 500 mg/50 mL vial ofrituximab.

In some embodiments, the kit comprises two 375 mg doses of Compound 1and one 10 mg/mL vial of rituximab. In some embodiments, the kitcomprises two 375 mg doses of Compound 1 and one 100 mg/10 mL vial ofrituximab. In some embodiments, the kit comprises two 375 mg doses ofCompound 1 and one 500 mg/50 mL vial of rituximab. In some embodiments,the kit comprises two 500 mg doses of Compound 1 and one 10 mg/mL doseof rituximab. In some embodiments, the kit comprises two 500 mg doses ofCompound 1 and one 100 mg/10 mL vial of rituximab. In some embodiments,the kit comprises two 500 mg doses of Compound 1 and one 500 mg/50 mLvial of rituximab.

IV. Formulations Comprising Compound 1

As described above, provided methods comprise administering to a patientin need thereof a pharmaceutically acceptable composition comprisingCompound 1, wherein the pharmaceutically acceptable composition is anoral dosage form. In some embodiments, the pharmaceutically acceptablecomposition is formulated as a capsule.

In certain embodiments, provided methods comprise administering to apatient in need thereof a pharmaceutically acceptable composition whichcomprises Compound 1, and one or more pharmaceutically acceptableexcipients, such as, for example, binders, film coatings, diluents,disintegrants, surfactants (wetting agents), lubricants and glidants(adsorbents), or combinations thereof. One skilled in the art willreadily appreciate that the category under which a particular componentis listed is not intended to be limiting; in some cases a particularcomponent might appropriately fit in more than one category. Also, aswill be appreciated, the same component can sometimes perform differentfunctions, or can perform more than one function, in the context of aparticular formulation, for example depending upon the amount of theingredient and/or the presence of other ingredients and/or activecompound(s). In some embodiments, the pharmaceutically acceptablecomposition is a blended powder.

i. Binders and Diluents

Pharmaceutical compositions for use in the present invention maycomprise one or more binders. Binders are used in the formulation ofsolid oral dosage forms to hold the active pharmaceutical ingredient andinactive ingredients together in a cohesive mix. In some embodiments,pharmaceutical compositions of the present invention comprise about 5%to about 50% (w/w) of one or more binders and/or diluents. In someembodiments, pharmaceutical compositions of the present inventioncomprise about 20% (w/w) of one or more binders and/or diluents.Suitable binders and/or diluents (also referred to as “fillers”) areknown in the art. Representative binders and/or diluents include, butare not limited to, starches such as celluloses (low molecular weightHPC (hydroxypropyl cellulose), microcrystalline cellulose (e.g.,Avicel), low molecular weight HPMC (hydroxypropyl methylcellulose), lowmolecular weight carboxymethyl cellulose, ethylcellulose), sugars suchas lactose (i.e. lactose monohydrate), sucrose, dextrose, fructose,maltose, glucose, and polyols such as sorbitol, mannitol, lactitol,malitol and xylitol, or a combination thereof. In some embodiments, aprovided composition comprises a binder of microcrystalline celluloseand/or lactose monohydrate.

ii. Disintegrants

Pharmaceutical compositions for use in the present invention may furthercomprise one or more disintegrants. Suitable disintegrants are known inthe art and include, but are not limited to, agar, calcium carbonate,sodium carbonate, sodium bicarbonate, cross-linked sodium carboxymethylcellulose (croscarmellose sodium), sodium carboxymethyl starch (sodiumstarch glycolate), microcrystalline cellulose, or a combination thereof.In some embodiments, provided formulations comprise from about 1%, toabout 25% disintegrant, based upon total weight of the formulation.

iii. Surfactants

Surfactants, also referred to as bioavailability enhancers, are wellknown in the art and typically facilitate drug release and absorption byenhancing the solubility of poorly-soluble drugs. Representativesurfactants include, but are not limited to, poloxamers, polyoxyethyleneethers, polyoxyethylene fatty acid esters, polyethylene glycol fattyacid esters, polyoxyethylene hydrogenated castor oil, polyoxyethylenealkyl ether, polysorbates, and combinations thereof. In certainembodiments, the surfactant is a poloxamer. In some such embodiments,the poloxamer is poloxamer 407. In some embodiments, compositions foruse in the present invention comprise from about 1% to about 30% byweight of surfactant, based upon total weight of the blended powder.

iv. Lubricants

Pharmaceutical compositions of the present invention may furthercomprise one or more lubricants. Lubricants are agents added in smallquantities to formulations to improve certain processingcharacteristics. Lubricants prevent the formulation mixture fromsticking to the compression machinery and enhance product flow byreducing interparticulate friction. Representative lubricants include,but are not limited to, magnesium stearate, glyceryl behenate, sodiumstearyl fumarate and fatty acids (i.e. palmitic and stearic acids). Incertain embodiments, a lubricant is magnesium stearate. In someembodiments, provided formulations comprise from about 0.2% to about 3%lubricant, based upon total weight of given formulation.

v. Glidants

Pharmaceutical compositions of the present invention may furthercomprise one or more glidants. Representative glidants include, but arenot limited to, silicas (i.e. fumed silica), microcrystallinecelluloses, starches (i.e. corn starch) and carbonates (i.e. calciumcarbonate and magnesium carbonate). In some embodiments, providedformulations comprise from about 0.2% to about 3% glidant, based upontotal weight of given formulation.

vi.N-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamidebesylate

As described above, the present invention provides a method of treatinga disease or disorder selected from chronic lymphocytic leukemia andsmall lymphocytic lymphoma, the method comprising administering to apatient in need thereof Compound 1 in combination with rituximab. Thebesylate salt of Compound 1,N-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamidebenzenesulfonic acid salt, has recently been identified and is currentlyin clinical trials as monotherapy in subjects with relapsed orrefractory B-cell non-Hodgkin's lymphoma (B-NHL), chronic lymphocyticleukemia (CLL) and Waldenstrom's macroglobulinemia (WM). Thus, in someembodiments, provided methods comprise administering to a patient inneed thereof a besylate salt of Compound 1.

In some embodiments, provided methods comprise administering to apatient in need thereof a pharmaceutically acceptable compositioncomprising from about 5% to about 60% of the besylate salt of Compound1, based upon total weight of the formulation. In some embodiments,provided methods comprise administering to a patient in need thereof apharmaceutically acceptable composition comprising from about 5% toabout 15% or about 7% to about 15% or about 7% to about 10% or about 9%to about 12% of the besylate salt of Compound 1, based upon total weightof the composition. In some embodiments, provided methods compriseadministering to a patient in need thereof a pharmaceutically acceptablecomposition comprising from about 25% to about 75% or about 30% to about60% or about 40% to about 50% or about 40% to about 45% of the besylatesalt of Compound 1, based upon total weight of the formulation. Incertain embodiments, provided methods comprise administering to apatient in need thereof a pharmaceutically acceptable compositioncomprising from about 8%, about 9%, about 10%, about 11%, about 12%,about 13%, about 20%, about 30%, about 40%, about 41%, about 42%, about43%, about 44%, about 45%, about 50%, about 60%, about 70%, or about 75%of the besylate salt of Compound 1, based upon total weight of givencomposition or formulation.

In some such embodiments, provided methods comprise administering to apatient in need thereof a pharmaceutical composition comprising a unitdose of Compound 1, wherein Compound 1 is in the form of a besylatesalt. In some such embodiments, the unit dose is an amount sufficient toprovide about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 125mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg or about 250mg of the free base of Compound 1. In some embodiments, thepharmaceutical composition comprising the besylate salt of Compound 1 isa solid oral dosage form.

In some embodiments, the present invention provides a method oftreating, stabilizing or lessening the severity or progression of adisease or disorder selected from the group consisting of chroniclymphocytic leukemia and small lymphocytic lymphoma, the methodcomprising administering to a patient in need thereof Compound 1 incombination with rituximab, wherein Compound 1 is administered as thebesylate salt. In some such embodiments, the besylate salt of Compound 1is administered in the form of a composition comprising one or morepharmaceutically acceptable excipients selected from binders, filmcoatings, diluents, disintegrants, surfactants, lubricants and glidants.In some embodiments, the present invention provides a method oftreating, stabilizing or lessening the severity or progression of adisease or disorder selected from the group consisting of chroniclymphocytic leukemia and small lymphocytic lymphoma, the methodcomprising administering to a patient in need thereof each of Compound1, rituximab, fludarabine and cyclophosphamide, wherein Compound 1 isadministered as the besylate salt. In some such embodiments, thebesylate salt of Compound 1 is administered in the form of a compositioncomprising one or more pharmaceutically acceptable excipients selectedfrom binders, film coatings, diluents, disintegrants, surfactants,lubricants and glidants. In some embodiments, the present inventionprovides a method of treating, stabilizing or lessening the severity orprogression of a disease or disorder selected from the group consistingof chronic lymphocytic leukemia and small lymphocytic lymphoma, themethod comprising administering to a patient in need thereof each ofCompound 1, rituximab and bendamustine, wherein Compound 1 isadministered as the besylate salt. In some such embodiments, thebesylate salt of Compound 1 is administered in the form of a compositioncomprising one or more pharmaceutically acceptable excipients selectedfrom binders, film coatings, diluents, disintegrants, surfactants,lubricants and glidants.

In some embodiments, the present invention provides a method oftreating, stabilizing or lessening the severity or progression of adisease or disorder selected from the group consisting of chroniclymphocytic leukemia and small lymphocytic lymphoma, the methodcomprising administering to a patient in need thereof a pharmaceuticalcomposition comprising the besylate salt of Compound 1 in combinationwith rituximab, wherein the amount of besylate salt of Compound 1 issufficient to deliver about 125 mg, about 250 mg, about 325 mg, about375 mg, about 400 mg, about 500 mg, about 625 mg, about 750 mg, about1000 mg or about 1250 mg of the free base of Compound 1. In some suchembodiments, the pharmaceutical composition further comprises one ormore pharmaceutically acceptable excipients selected from binders, filmcoating, diluents, disintegrants, surfactants, lubricants and glidants.In some such embodiments, the pharmaceutical composition comprises oneor more pharmaceutically acceptable excipients selected frommicrocrystalline cellulose, lactose monohydrate, sodium starch,poloxamer 407, fumed silica and magnesium stearate. In some embodiments,the present invention provides a method of treating, stabilizing orlessening the severity or progression of a disease or disorder selectedfrom the group consisting of chronic lymphocytic leukemia and smalllymphocytic lymphoma, the method comprising administering to a patientin need thereof a pharmaceutical compositions comprising each of thebesylate salt of Compound 1 (i.e., Compound1 besylate), rituximab,fludarabine and cyclophosphamide, wherein the amount of besylate salt ofCompound 1 is sufficient to deliver about 125 mg, about 250 mg, about325 mg, about 375 mg, about 400 mg, about 500 mg, about 625 mg, about750 mg, about 1000 mg or about 1250 mg of the free base of Compound 1.In some embodiments, the present invention provides a method oftreating, stabilizing or lessening the severity or progression of adisease or disorder selected from the group consisting of chroniclymphocytic leukemia and small lymphocytic lymphoma, the methodcomprising administering to a patient in need thereof a pharmaceuticalcompositions comprising each of the besylate salt of Compound 1 (i.e.,Compound1 besylate), rituximab and bendamustine, wherein the amount ofbesylate salt of Compound 1 is sufficient to deliver about 125 mg, about250 mg, about 325 mg, about 375 mg, about 400 mg, about 500 mg, about625 mg, about 750 mg, about 1000 mg or about 1250 mg of the free base ofCompound 1.

V. Process for Preparing Pharmaceutical Compositions Comprising Compound1 Dry Blend Process:

MilledN-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamidebesylate, milled microcrystalline cellulose, milled sodium starchglycolate, milled lactose monohydrate, milled poloxamer 407, and sievedfumed silica are weighed and mechanically blended. An intragranularportion of sieved magnesium stearate (2.0%, per Table 1, below) is addedto the blender and the formulation blended. This blended formulation isthen roller compacted, milled, and then blended. The blended formulationis additionally roller compacted, milled and then blended. The remainderor extragranular portion of the magnesium stearate (0.5%, per Table 1,below) is added and the final formulation is blended. Capsules areeither mechanically filled or manually filled via the flood fill method.

All features of each of the aspects of the invention apply to all otheraspects mutatis mutandis. Each of the references referred to herein,including but not limited to patents, patent applications and journalarticles, is incorporated by reference herein as though fully set forthin its entirety.

In order that the invention described herein may be more fullyunderstood, the following examples are set forth. It should beunderstood that these examples are for illustrative purposes only andare not to be construed as limiting this invention in any manner.

EXEMPLIFICATION Example 1 Dose Escalation Study

N-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamidebesylate is a chemically synthesized small molecule substitutedpyrimidine developed as the benzenesulfonic acid salt and is a white tooff-white crystalline powder.N-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamidebesylate is an oral, potent (IC₅₀<0.5 nM) and selective small moleculeinhibitor of Btk.N-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamidebesylate exhibits solubility of approximately 0.16 mg/mL in water and amaximum aqueous solubility of 0.40 mg/mL at approximately pH 3.0. Thesolubility ofN-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamidebesylate in ethanol is approximately 10 mg/mL.N-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamidebesylate exhibits no environmental instabilities (i.e. heat, acid, base)that require special handling.

N-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamidebesylate was formulated into capsules containing the components andquantities listed in Table 1 to obtain the study drug. The capsuleslisted in Table 1 will be administered during the dose escalation andexpansion cohort studies.

TABLE 1 Components of N-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin- 4-ylamino)phenyl)acrylamidebesylate capsules Amount per Component 125 mg Capsule Capsule shell 1,size 0 white capsule N-(3-(5-fluoro-2-(4-(2- 174.30 mgmethoxyethoxy)phenylamino) (125 mg free pyrimidin-4-ylamino)phenyl)base) acrylamide besylate Microcrystalline cellulose 101.68 mg Lactosemonohydrate 41.50 mg Sodium starch glycolate 41.50 mg Poloxamer 40741.50 mg Fumed silica 4.15 mg Magnesium stearate 10.38 mg^(‡) ^(‡)2.0%(8.30 mg) intragranular; 0.5% (2.08 mg) extragranular.

Rituximab is provided to the physician/investigator in 10 mg/mL vialscomprising 100 mg/10 mL or 500 mg/50 mL. Prior to administration,rituximab is diluted to a dose of 1 mg/mL, 2 mg/mL, 3 mg/mL or 4 mg/mLwith either 5% dextrose in water or 0.9% sodium chloride. Rituximab isthereafter administered as a 1 mg/mL to 4 mg/mL infusion according tothe dosages set forth in Table 2, below.

Study Design

Subjects with relapsed or refractory CLL or SLL who failed at least oneprior treatment regimen were enrolled in a “3+3” dose escalation andexpansion study to determine the Not Tolerated Dose (NTD), the OptimalBiologic Effect dose (OBE) and the Maximum Tolerated Dose (MTD) of thecombination of Compound 1 and rituximab. Approximately 30-42 patientsare expected to be enrolled in the study.

Study treatment was administered in 28-day cycles at specified doselevels as scheduled until disease progression, unacceptable toxicity, ordiscontinuation for any other reason. Subjects will continue on thestarting dose until the preliminary recommended Phase 2 dose (RP2D) isdetermined, at which point they can be switched to the preliminary RP2D.

N-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamidebesylate and rituximab were administered according to the cohorts listedin Table 2:

TABLE 2 Study Dosing Schema for Escalating Dose Portion of StudyN-(3-(5-fluoro-2-(4-(2- methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl) COHORT acrylamide besylate Rituximab 1 375mg BID 6 doses administered 2 500 mg BID as follows: 375 mg/m² on cycle1 day 2 500 mg/m² on cycles 2-6 day 1

Within each cohort, subjects were treated PO (oral) BID (daily) withN-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamide besylate during two initial 28-day treatment cycles and wereassessed for safety, tolerability and DLT, as well as pharmacokinetic(“PK”), pharmacodynamic (“PD”), and disease response. In certaininstances, the physician-investigator may elect to rest a patient duringthe study, during which time the patient does not receive treatment. Forexample, the physician-investigator may elect to rest a patient due tooccurrence or recurrence of adverse events. For purposes of clarity, apatient who has been rested is still enrolled in the study until thephysician-investigator determines that the patient should not continuetreatment, at which time such patients are discontinued from furthertreatment. In this context, treatment duration refers to the time apatient is enrolled in the study, inclusive of all rest periods, untiltreatment is discontinued.

Rituximab was administered as a single intravenous (IV) infusion. Theinitial infusion during cycle 1 was administered at 375 mg/m²;subsequent infusions during cycles 2 through 6 were administered at 500mg/m². Administration of rituximab began on day 2 of cycle 1 and on day1 of each cycle thereafter. Following the cycle 6 infusion, rituximabwill be discontinued. Subjects may continue on treatment withN-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamidebesylate if appropriate. The first infusion of rituximab was at a rateof 50 mg/hr. In the absence of infusion toxicity, the infusion rate willbe increased by 50 mg/hr increments every 30 minutes, to a maximum of400 mg/hr. Each subsequent infusion will be initiated at 100 mg/hr. Inthe absence of infusion toxicity, the infusion rate will be increased by100 mg/hr increments at 30 minute intervals to a maximum of 400 mg/hr.

The dose level at which a patient is enrolled will be based on whichcohort is open at the time of enrollment. Dose escalation, viaenrollment in the next higher dose, is allowed only if none (0) of thefirst three (3) subjects enrolled in any cohort experience dose limitingtoxicity (DLT). If one (1) of the first three (3) subjects dosed in anycohort experiences a DLT in cycle 1, three (3) more subjects will beenrolled in that dose cohort. A dose level will be considered to bebelow the NTD if <1 of 3 DLT evaluable subjects enrolled experiences aDLT during the first 2 cycles. A dose will be considered a NTD when atleast two (2) of six (6) DLT-evaluable subjects in that cohortexperience a DLT. A MTD will be declared when at least six (6) subjectshave been enrolled and safely complete cycle 1 at that dose level. TheMTD is defined as the last dose below the NTD with zero (0) or one (1)DLT-evaluable subject experiencing DLT during the first two 28-daycycles.

During the dose escalation phase, a decision to enroll the next higherdose cohort will be based on review of safety and DLT-evaluablepatients. The OBE dose is defined as follows:

-   -   a reduction of ≧50% in the size of lymph nodes in ≧two (2) of        six (6) subjects; and/or    -   no further increase in exposure with increasing doses; and/or    -   a ≧25% increase in lymphocytosis in four (4) of six (6) subjects        during the first three 28-day cycles not assessed as progressive        disease.

Results. “Complete Response” (CR) is defined per IWCLL criteria, 2008.(No LN>1.5 cm, no hepatomegaly, splenomegaly, ALC <4000/uL,normocellular marrow <30% lymphocytes, ANC>1500, Platelet Count>100,000and Hgb>11.0 g/dL). “Partial Response” (PR) was assessed via IWCLLguidelines (at least 2 of the following criteria—lymph node (LN)decrease ≧50%; hepatomegaly decrease ≧50%; splenomegaly decrease ≧50%;ALC decrease ≧50%; and at least 1 of the following—plateletcount >100,000; ANC>1500/uL or Hgb>11.0 g/dL). A status of PR is theinvestigator's assessment based on a physical exam evaluation of lymphnodes, spleen and liver and laboratory values of blood counts. ConfirmedPR also includes imaging of tumor lesions by CT scan.

Three subjects were enrolled in cohort 1 and treated withN-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamide besylate in combination with rituximab. Of the three subjectsenrolled in cohort 1 on Oct. 16, 2013, one subject is currently in theninth 28-day cycle and two subjects are currently in the eighth 28-daycycle. Because none of these subjects experienced a DLT during the first28-day cycle, three subjects were enrolled in cohort 2. Two of the 3subjects in cohort 2 completed the first two 28 day cycles without aDLT. The third subject has experienced a DLT of grade 3 fatigue andweakness as graded by the NCI-CTCAE guidelines. Three more subjects wereenrolled into cohort 2 for further assessment of DLTs and the safety ofthis dose level.

FIG. 1 summarizes the enrollment and response assessment of patients.All three patients in Cohort 1 have been assessed as having a partialresponse to study treatment. In all three cases the investigators'assessment of PR as of cycle 3 has been confirmed by CT scans showing agreater than 50% reduction in the size of lymph nodes compared topre-treatment baseline values. The first subject enrolled is currentlybeing treated in cycle 9 and continues to maintain a partial response.The other two patients in cohorts 1 have continued with treatment intocycle 8 and also maintain a partial response to treatment. Of the sixsubjects treated in cohort 2, two subjects have achieved a partialresponse by the start of cycle 3 according to the investigatorassessment by examination. One subject has exhibited a partial responseby cycle 5 as per investigator assessment. Response assessments for theadditional 3 subjects enrolled in cohort 2 is pending.

After full enrollment of each dose escalation cohort and completion ofthe second cycle of treatment for each dose escalation cohort, thenumber and type of DLTs and adverse events (AE) occurring during thefirst two cycles will be evaluated. Subjects will remain on study untilthe subject discontinues due to disease progression, unacceptabletoxicity, withdrawal of consent or any other reason determined by thephysician. Preliminary evidence of efficacy will be evaluated.

Expansion Cohorts.

After completion of observation for DLTs in the dose escalation study,the accumulated safety, PK, and PD data will be evaluated to select apreliminary RP2D. The preliminary RP2D will be evaluated in expandedcohorts of 24 subjects or a more complete safety profile and furtherpreliminary evaluation of efficacy. If less than 9 of 24 subjectsexperience DLTs, then this dose level will be declared the RP2D to beused in further studies. If DLTs are experienced in greater than orequal to 9 of 24 subjects, this dose will be considered to have exceededthe MTD and the previous highest tolerated dose found in the doseescalation cohort of the study will be evaluated in 24 subjects. Thedose level will continue to be reduced in a stepwise fashion until lessthan 9 of 24 subjects experience DLTs.

During the expansion cohort, rituximab will be administered according tothe schedule set forth in the dose escalation cohorts. Rituximab will beadministered as a single intravenous (IV) infusion. The initial infusionduring cycle 1 will be administered at 375 mg/m²; subsequent infusionsduring cycles 2 through 6 will be administered at 500 mg/m².Administration of rituximab will begin on day 2 of cycle 1 and on day 1of each cycle thereafter. Following the cycle 6 infusion, rituximab willbe discontinued. Each subject will continue on treatment withN-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamidebesylate until disease progression, unacceptable toxicity or treatmentdiscontinuation for any other reason.

In certain instances, the physician-investigator may elect to rest apatient during the study, during which time the patient does not receivetreatment. For example, the physician-investigator may elect to rest apatient due to occurrence or recurrence of adverse events. For purposesof clarity, a patient who has been rested is still enrolled in the studyuntil the physician-investigator determines that the patient should notcontinue treatment, at which time such patients are discontinued fromfurther treatment. In this context, treatment duration refers to thetime a patient is enrolled in the study, inclusive of all rest periods,until treatment is discontinued.

Adverse Events.

For all cohorts, dose limiting toxicities (DLTs) are defined asspecified adverse events (AEs) that are observed within the first two28-day cycles (approximately 56 days) and deemed to be related totreatment. Hematologic DLTs include Grade 4 anemia (hemoglobin decrease)or thrombocytopenia by NCI-CTCAE (v. 4.03) or by IWCLL criteria,whichever results in the lower blood threshold; Grade 4 neutropeniagreater than 5 days despite granulocyte colony-stimulating factor(G-CSF) support; and Grade 3 or higher febrile neutropenia.Lymphocytosis may be observed as a consequence of disease progressionbut has also been described as a redistribution (lymphocytle migrationand trafficking) phenomenon in subjects receiving another BTK inhibitoreven as lymph node disease responds to treatment. Therefore,lymphocytosis will not be rated for DLT. Reduction of malignantlymphocytosis is an intended therapeutic effect of treatment and willnot be considered for DLT.

Non-hematologic DLTs include Grade 4 or higher non-hematologic AEs ofany duration; Grade 3 total bilirubin elevation, whether symptomatic orasymptomatic; and any Grade 3 non-hematologic toxicity except nausea,vomiting and diarrhea lasting less than 24 hours following medicaltherapy; tumor lysis syndrome which does not progress to Grade 4 andresolves in less than 7 days with medical management is not considered aDLT; and transient, and Grade 3 non-hematologic laboratory anomaly thatis asymptomatic and rapidly reversible (returns to baseline or ≦Grade 1within 7 days) will not be considered a DLT.

Subjects without disease progression and without DLT at the end of thefirst two 28-day treatment cycles are eligible to continue receivingN-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamidebesylate in combination with rituximab for additional 28-day cyclesuntil (i) the patient experiences unacceptable toxicity, (ii) theunderlying malignancy progresses, (iii) the patient withdraws consent,or (iv) the treating physician-investigator otherwise determines thatthe patient should not continue treatment. Subjects experiencing a DLTmay remain on study treatment if the treating investigator determinesthat the subject is receiving a clinical benefit from the studytreatment. Rituximab will only be administered for the first 6 cycles;however patients continuing to benefit fromN-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamidebesylate can remain on treatment.

One subject in cohort 1 experienced herpetic esophagitis during cycle 2and two SAEs during cycle 5; an incidence of atrial fibrillation andpneumonia and an incidence of hypotension, altered mental status andpneumonia, both reports which were considered unrelated to the studydrugs. All 3 of these SAE reports required brief hospitalizations anddrug interruptions, however following re-challenged with drug thissubject has subsequently achieved further improvement in disease status.In cohort 2, one subject experienced scrotal abscess during cycle 2,which was deemed unrelated to the study drugs. Another cohort 2 subjectexperienced a Grade 3 fatigue during cycle 2 which was declared a DLT;however, the AE was not reported as serious. This patient remains onstudy treatment at a reduced dose ofN-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamidebesylate. One additional subject in cohort 2 experienced visualdisturbance during cycle 2, which was present at baseline. Two subjects,one from each cohort, reported Grade 3 neutropenia.

Example 2

One particular irreversible BTK inhibitor, Compound 2, was screenedagainst 342 kinases to ascertain kinase activity and/or selectivity:

The binding assay system for profiling kinase activity were based uponHotSpot technology (Reaction Biology Corp.; Malvern, Pa., USA) andutilized radio-isotope-based P81 filtration. Compound 2 was dissolved inpure DMSO to make a 10 mM stock solution and serial dilutions wereperformed to a final 3 μM test concentration. Substrates for the variouskinases tested against Compound 2 (substrate information available onthe Reaction Biology Corp. website) were prepared fresh daily inReaction Buffer. Any required cofactors were then added to the substratesolution. The identification and selection of the appropriate cofactorfor each kinase is within the ability of a person skilled in the art.See, for example, Handbook of Assay Development in Drug Discovery, Ed.Lisa K. Minor, 2006: CRC Press, Boca Raton, Fla.; Gao et al., “A broadactivity screen in support of a chemogenomic map for kinase signallingresearch and drug discovery,” Biochem J. 2013, 451(2): 313-28; and Eglenet al., “Drug discovery and the human kinome: Recent trends,”Pharmacology & Therapeutics 2011, 130(2): 144-156. The kinase was thenadded to the substrate solution and gently mixed. Compound 2 (5 nL) wasthen added to the kinase reaction mixture by acoustical droplet ejectionand preincubated for 30 min at room temperature. ³³P-ATP (100 μM) wasdelivered into the reaction mixture to initiate the reaction. This wasfollowed by incubation at room temperature for 2h. The reaction wasterminated and any unreacted phosphate was washed away using 0.1%phosphoric acid prior to detection utilizing a proprietary technology.The study was performed in duplicate and staurosporine, a non-selective,ATP-competitive kinase inhibitor, was used a the positive control in a10-dose IC50 mode with 3-fold serial dilutions starting at 1 μM, 50 μMor 100 μM. DMSO was used as the negative control.

Determination of Percent Inhibition.

Percent inhibition of a kinase by a test compound, e.g., Compound 2, wasdetermined according to the following formula: percentinhibition=[(kinase activity of negative control)−(kinase activity inpresence of a test compound, e.g., Compound 2)/(kinase activity ofnegative control)]×100. Percent inhibition was expressed as an averagewhere the assay was performed more than once, e.g., in duplicate.

Table 3 sets forth the average percent inhibition for Compound 2 againstvarious kinases:

TABLE 3 Average Percent Inhibition of Kinases by Compound 2 % EnzymeActivity (relative to DMSO controls) Average % Kinase Run 1 Run 2Inhibition ABL1 45.37 45.23 54.70 ABL2/ARG 37.26 37.07 62.84 ACK1 22.5922.50 77.46 AKT1 95.13 93.13 5.87 AKT2 93.65 97.89 4.23 AKT3 110.81102.89 −6.85 ALK 82.52 83.61 16.94 ALK1/ACVRL1 124.78 123.91 −24.35ALK2/ACVR1 385.44 385.47 −285.46 ALK3/BMPR1A 104.30 104.51 −4.41ALK4/ACVR1B 140.52 143.80 −42.16 ALK5/TGFBR1 106.06 110.00 −8.03ALK6/BMPR1B 176.94 183.87 −80.41 ARAF 102.92 100.02 −1.47 ARK5/NUAK113.26 13.31 86.72 ASK1/MAP3K5 100.91 102.32 −1.62 Aurora A 12.26 11.0388.35 Aurora B 13.18 12.94 86.94 Aurora C 26.24 29.79 71.98 AXL 48.5848.79 51.31 BLK 6.30 6.37 93.66 BMPR2 83.77 83.85 16.19 BMX/ETK 0.931.23 98.92 BRAF 100.83 99.10 0.04 BRK 25.10 24.97 74.96 BRSK1 87.1089.40 11.75 BRSK2 90.99 91.69 8.66 BTK 3.69 5.40 95.45 CAMK1a 109.10111.65 −10.38 CAMK1b 103.91 104.95 −4.43 CAMK1d 103.60 103.13 −3.36CAMK1g 99.95 101.49 −0.72 CAMK2a 109.14 110.06 −9.60 CAMK2b 96.40 95.813.89 CAMK2d 123.75 122.81 −23.28 CAMK2g 103.51 110.85 −7.18 CAMK4 102.16102.35 −2.26 CAMKK1 93.48 87.89 9.31 CAMKK2 74.65 71.82 26.77 CDC7/DBF4100.26 103.78 −2.02 CDK1/cyclin A 111.93 124.07 −18.00 CDK1/cyclin B87.16 87.03 12.91 CDK1/cyclin E 100.38 102.23 −1.30 CDK16/cyclin Y104.10 103.36 −3.73 CDK2/cyclin A 85.83 86.86 13.66 CDK2/cyclin A1 78.2577.74 22.00 CDK2/cyclin E 87.92 92.03 10.02 CDK3/cyclin E 78.15 76.9522.45 CDK4/cyclin D1 95.58 96.28 4.07 CDK4/cyclin D3 95.54 94.15 5.15CDK5/p25 89.55 92.26 9.10 CDK5/p35 94.97 96.06 4.49 CDK6/cyclin D1101.41 100.42 −0.92 CDK6/cyclin D3 99.55 100.13 0.16 CDK7/cyclin H 98.1097.18 2.36 CDK9/cyclin K 81.70 82.16 18.07 CDK9/cyclin T1 87.76 91.5410.35 CHK1 93.14 94.56 6.15 CHK2 25.85 25.28 74.43 CK1a1 107.76 105.57−6.67 CK1d 99.87 100.20 −0.03 CK1epsilon 101.51 102.41 −1.96 CK1g1 88.7790.27 10.48 CK1g2 89.13 85.74 12.57 CK1g3 84.15 85.84 15.01 CK2a 117.05123.24 −20.15 CK2a2 98.25 105.38 −1.81 c-Kit 77.47 79.45 21.54 CLK174.39 77.44 24.08 CLK2 50.21 50.65 49.57 CLK3 90.36 95.83 6.90 CLK457.40 53.92 44.34 c-MER 66.48 66.40 33.56 c-MET 102.62 100.36 −1.49COT1/MAP3K8 101.72 100.94 −1.33 CSK 81.71 81.81 18.24 c-Src 28.17 27.8871.97 CTK/MATK 102.20 103.87 −3.03 DAPK1 102.79 93.48 1.86 DAPK2 108.05111.72 −9.89 DCAMKL1 98.43 97.52 2.02 DCAMKL2 100.15 99.50 0.18 DDR125.70 24.12 75.09 DDR2 102.90 104.85 −3.87 DLK/MAP3K12 74.17 80.18 22.82DMPK 104.46 102.05 −3.25 DMPK2 97.36 99.56 1.54 DRAK1/STK17A 82.93 80.5718.25 DYRK1/DYRK1A 87.66 88.41 11.96 DYRK1B 78.60 80.92 20.24 DYRK260.98 62.12 38.45 DYRK3 85.99 85.89 14.06 DYRK4 105.18 105.53 −5.35 EGFR19.23 20.17 80.30 EPHA1 99.47 101.01 −0.24 EPHA2 84.23 84.12 15.82 EPHA396.21 100.18 1.81 EPHA4 92.86 88.96 9.09 EPHA5 89.11 93.57 8.66 EPHA695.52 102.01 1.24 EPHA7 60.01 64.14 37.93 EPHA8 94.25 93.79 5.98 EPHB179.45 79.66 20.44 EPHB2 104.27 106.64 −5.45 EPHB3 99.17 98.21 1.31 EPHB481.25 81.45 18.65 ERBB2/HER2 39.81 37.50 61.34 ERBB4/HER4 9.15 8.3291.27 ERK1 97.44 99.93 1.32 ERK2/MAPK1 105.72 102.79 −4.25 ERK5/MAPK7100.83 99.91 −0.37 ERK7/MAPK15 63.77 66.05 35.09 FAK/PTK2 62.84 62.8637.15 FER 88.07 88.26 11.83 FES/FPS 63.95 65.95 35.05 FGFR1 41.38 39.3159.66 FGFR2 37.28 35.97 63.37 FGFR3 32.14 31.78 68.04 FGFR4 66.32 63.4435.12 FGR 39.73 40.47 59.90 FLT1/VEGFR1 83.62 79.20 18.59 FLT3 3.19 3.2596.78 FLT4/VEGFR3 43.82 44.24 55.97 FMS 64.50 67.34 34.08 FRK/PTK5100.54 97.96 0.75 FYN 81.08 81.84 18.54 GCK/MAP4K2 100.27 100.02 −0.14GLK/MAP4K3 102.03 108.13 −5.08 GRK1 103.55 103.40 −3.47 GRK2 104.15103.91 −4.03 GRK3 99.45 100.95 −0.20 GRK4 107.15 106.07 −6.61 GRK5103.03 102.15 −2.59 GRK6 102.34 103.73 −3.03 GRK7 89.14 90.93 9.96 GSK3a75.97 74.98 24.53 GSK3b 121.82 122.56 −22.19 Haspin 90.56 89.91 9.77 HCK71.99 68.97 29.52 HGK/MAP4K4 98.87 98.60 1.27 HIPK1 87.63 91.73 10.32HIPK2 95.75 98.61 2.82 HIPK3 112.11 118.15 −15.13 HIPK4 93.61 93.78 6.31HPK1/MAP4K1 85.77 88.42 12.90 IGF1R 82.83 85.99 15.59 IKKa/CHUK 91.4391.50 8.54 IKKb/IKBKB 95.44 97.06 3.75 IKKe/IKBKE 78.26 78.18 21.78 IR84.52 84.68 15.40 IRAK1 76.49 74.41 24.55 IRAK4 88.61 86.52 12.43IRR/INSRR 88.90 90.97 10.07 ITK 7.97 7.95 92.04 JAK1 59.70 59.03 40.63JAK2 108.64 114.63 −11.63 JAK3 2.53 2.73 97.37 JNK1 88.14 87.66 12.10JNK2 92.48 95.08 6.22 JNK3 110.08 115.56 −12.82 KDR/VEGFR2 83.35 81.2417.70 KHS/MAP4K5 94.89 90.82 7.15 LATS1 89.52 89.80 10.34 LATS2 88.4091.16 10.22 LCK 64.67 63.36 35.99 LCK2/ICK 100.94 95.10 1.98 LIMK1 60.9861.36 38.83 LIMK2 101.00 100.73 −0.86 LKB1 99.24 95.94 2.41 LOK/STK1043.10 43.01 56.94 LRRK2 33.02 35.12 65.93 LYN 71.67 73.24 27.54 LYN B92.90 95.69 5.70 MAPKAPK2 109.81 105.91 −7.86 MAPKAPK3 102.28 102.63−2.45 MAPKAPK5/PRAK 109.84 113.33 −11.58 MARK1 90.93 97.35 5.86MARK2/PAR-1Ba 99.58 97.83 1.30 MARK3 101.48 100.37 −0.92 MARK4 82.1780.87 18.48 MEK1 109.05 111.60 −10.33 MEK2 106.36 104.95 −5.66 MEK3117.30 113.34 −15.32 MEKK1 112.92 116.55 −14.74 MEKK2 108.13 113.65−10.89 MEKK3 101.68 106.39 −4.03 MELK 108.80 107.34 −8.07 MINK/MINK1100.72 97.49 0.90 MKK4 116.84 116.35 −16.60 MKK6 96.36 97.41 3.11MLCK/MYLK 95.57 95.24 4.59 MLCK2/MYLK2 71.62 68.13 30.13 MLK1/MAP3K914.50 14.05 85.72 MLK2/MAP3K10 45.39 45.33 54.64 MLK3/MAP3K11 27.8925.23 73.44 MNK1 97.78 94.26 3.98 MNK2 83.04 83.21 16.88 MRCKa/CDC42BPA113.51 115.15 −14.33 MRCKb/CDC42BPB 106.78 105.47 −6.12 MSK1/RPS6KA594.87 99.72 2.70 MSK2/RPS6KA4 103.35 97.38 −0.36 MSSK1/STK23 108.42104.81 −6.62 MST1/STK4 70.20 68.65 30.58 MST2/STK3 91.08 88.64 10.14MST3/STK24 86.56 84.46 14.49 MST4 97.85 104.33 −1.09 MUSK 70.16 68.0530.89 MYLK3 113.48 116.49 −14.98 MYO3b 103.52 101.67 −2.59 NEK1 52.7453.95 46.65 NEK11 92.53 92.84 7.32 NEK2 102.22 106.65 −4.44 NEK3 76.6575.11 24.12 NEK4 84.64 88.33 13.52 NEK5 60.25 60.87 39.44 NEK6 105.95105.43 −5.69 NEK7 98.29 100.34 0.68 NEK9 83.83 84.30 15.94 NLK 99.61102.97 −1.29 OSR1/OXSR1 116.94 121.56 −19.25 P38a/MAPK14 105.80 107.56−6.68 P38b/MAPK11 101.64 101.17 −1.41 P38d/MAPK13 98.94 99.78 0.64 P38g105.35 105.71 −5.53 P70S6K/RPS6KB1 85.89 81.02 16.54 P70S6Kb/RPS6KB295.60 96.10 4.15 PAK1 96.27 98.58 2.58 PAK2 95.53 94.87 4.80 PAK3 91.8695.22 6.46 PAK4 98.70 96.40 2.45 PAK5 122.54 132.33 −27.43 PAK6 79.3084.76 17.97 PASK 94.65 93.08 6.14 PBK/TOPK 98.58 95.27 3.07 PDGFRa 63.9362.78 36.65 PDGFRb 47.74 47.41 52.42 PDK1/PDPK1 106.60 105.33 −5.96PHKg1 85.80 85.18 14.51 PHKg2 117.59 111.88 −14.73 PIM1 103.69 103.94−3.82 PIM2 135.59 129.41 −32.50 PIM3 103.69 99.97 −1.83 PKA 85.81 85.1314.53 PKAcb 49.70 51.07 49.62 PKAcg 127.92 127.73 −27.82 PKCa 88.4088.80 11.40 PKCb1 72.71 72.06 27.62 PKCb2 50.13 48.85 50.51 PKCd 100.4196.02 1.78 PKCepsilon 93.22 94.21 6.29 PKCeta 108.82 116.79 −12.81 PKCg83.88 84.16 15.98 PKCiota 105.24 105.70 −5.47 PKCmu/PRKD1 74.85 75.8524.65 PKCnu/PRKD3 80.06 79.79 20.07 PKCtheta 83.65 84.12 16.12 PKCzeta99.81 95.15 2.52 PKD2/PRKD2 86.37 86.22 13.70 PKG1a 87.99 94.55 8.73PKG1b 85.36 87.08 13.78 PKG2/PRKG2 87.22 84.45 14.16 PKN1/PRK1 93.8392.30 6.94 PKN2/PRK2 93.30 91.42 7.64 PKN3/PRK3 106.24 108.40 −7.32 PLK191.11 92.77 8.06 PLK2 86.63 86.74 13.32 PLK3 96.95 100.49 1.28 PLK4/SAK54.75 55.16 45.04 PRKX 97.87 98.53 1.80 PYK2 70.59 70.13 29.64 RAF183.34 83.62 16.52 RET 13.17 13.63 86.60 RIPK2 77.59 75.01 23.70 RIPK3116.80 120.69 −18.75 RIPK5 96.13 99.22 2.32 ROCK1 107.64 105.14 −6.39ROCK2 102.29 101.47 −1.88 RON/MST1R 103.11 101.37 −2.24 ROS/ROS1 13.5213.26 86.61 RSK1 73.29 72.45 27.13 RSK2 82.57 84.26 16.59 RSK3 85.8085.16 14.52 RSK4 77.21 77.21 22.79 SGK1 99.71 99.55 0.37 SGK2 71.3876.59 26.02 SGK3/SGKL 99.54 105.31 −2.42 SIK1 48.40 48.72 51.44 SIK256.26 57.07 43.34 SIK3 91.33 92.80 7.93 SLK/STK2 74.89 75.27 24.92SNARK/NUAK2 83.70 84.92 15.69 SRMS 123.69 122.52 −23.11 SRPK1 98.8196.73 2.23 SRPK2 90.92 89.61 9.73 SSTK/TSSK6 107.37 99.84 −3.60 STK1622.93 21.04 78.01 STK22D/TSSK1 88.04 89.72 11.12 STK25/YSK1 94.47 94.105.72 STK32B/YANK2 95.81 93.60 5.30 STK32C/YANK3 104.94 107.08 −6.01STK33 51.19 52.54 48.14 STK38/NDR1 92.71 93.70 6.79 STK38L/NDR2 106.6895.97 −1.33 STK39/STLK3 91.63 92.89 7.74 SYK 78.04 77.10 22.43 TAK173.39 71.98 27.32 TAOK1 100.50 96.25 1.63 TAOK2/TAO1 98.49 94.44 3.53TAOK3/JIK 94.52 90.00 7.74 TBK1 58.13 59.19 41.34 TEC 10.36 11.47 89.08TESK1 97.22 98.52 2.13 TGFBR2 98.07 102.31 −0.19 TIE2/TEK 102.88 107.55−5.21 TLK1 102.43 103.40 −2.91 TLK2 107.74 104.89 −6.32 TNIK 66.84 67.7732.69 TNK1 15.91 16.61 83.74 TRKA 117.40 117.94 −17.67 TRKB 86.53 85.0014.24 TRKC 35.08 32.34 66.29 TSSK2 97.01 96.27 3.36 TSSK3/STK22C 132.55132.59 −32.57 TTBK1 101.11 102.19 −1.65 TTBK2 102.05 99.67 −0.86 TXK0.42 0.03 99.77 TYK1/LTK 77.75 75.81 23.22 TYK2 48.20 46.63 52.58TYRO3/SKY 95.11 96.84 4.03 ULK1 100.12 101.06 −0.59 ULK2 102.27 109.21−5.74 ULK3 79.58 76.98 21.72 VRK1 84.28 89.86 12.93 VRK2 94.00 95.885.06 WEE1 73.06 74.89 26.03 WNK1 112.15 112.04 −12.10 WNK2 92.35 95.286.19 WNK3 91.35 91.97 8.34 YES/YES1 18.72 18.27 81.50 ZAK/MLTK 83.7185.00 15.65 ZAP70 110.35 108.07 −9.21 ZIPK/DAPK3 109.39 113.13 −11.26

Example 3

Capsules comprisingN-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamidebesylate (Compound 1 besylate) for use in this study correspond to Table1 in Example 1.

Rituximab is provided to the physician/investigator in 10 mg/mL vialscomprising 100 mg/10 mL or 500 mg/50 mL. Prior to administration,rituximab is diluted to a dose of 1 mg/mL, 2 mg/mL, 3 mg/mL or 4 mg/mLwith either 5% dextrose in water or 0.9% sodium chloride. Rituximab isthereafter administered as a 1 mg/mL to 4 mg/mL infusion according tothe dosages set forth in Table 4, below.

Fludarabine is approved as Fludara® and is commercially available in avial containing a sterile lyophilized solid cake which contains 50 mg offludarabine phosphate, 50 mg of mannitol, and sodium hydroxide. Thesolid cake is reconstituted with 2 mL of Sterile Water for InjectionUSP, resulting in a 25 mg/mL solution.

Cyclophosphamide is approved as Cytoxan® and is commerically availableas a sterile powder which con be reconstituted according to the packageinsert.

Bendamustine is approved as Treanda® and is commercially available as asa single-use vial containing 100 mg of bendamustine hydrochloride as alyophilized powder. The powder is reconstituted with 20 mL of SterileWater for Injection USP, resulting in a 5 mg/mL solution, which isfurther diluted with 0.9% Sodium Chloride Injection, USP or 2.5%Dextrose/0.45% Sodium Chloride Injection, USP immediately prior toinjection (final concentration of 0.2-0.6 mg/mL).

Study Design

Subjects with relapsed or refractory CLL or SLL who failed at least oneprior treatment regimen will be enrolled in a “3+3” dose escalation andexpansion study to determine the Not Tolerated Dose (NTD), the OptimalBiologic Effect dose (OBE) and the Maximum Tolerated Dose (MTD) ofCompound 1, rituximab, fludarabine and cyclophosphamide (Arm A) andCompound 1, rituximab and bendamustine (Arm B). Approximately 30-42patients are expected to be enrolled in the study.

Study treatment will be administered in 28-day cycles at specified doselevels as scheduled until disease progression, unacceptable toxicity, ordiscontinuation for any other reason. Subjects will continue on thestarting dose until the preliminary recommended Phase 2 dose (RP2D) isdetermined, at which point they can be switched to the preliminary RP2D.

Compound 1 besylate, rituximab, fludarabine and cyclophosphamide will beadministered according to the cohorts for Arm A of the study, listed inTable 4:

TABLE 4 Study Dosing Schema for Arm A Escalating Dose Portion of StudyCompound 1 Cyclo- COHORT besylate Rituximab Fludarabine phosphamide 1A375 mg BID: 6 doses 18 doses 18 doses days 8-28 administeredadministered administered for cycle 1 as follows: as follows: asfollows: days 1-28 375 mg/m² 25 mg/m² 250 mg/m² for on day 1 on days 1-3on days 1-3 subsequent for cycle 1 for cycles for cycles cycles 500mg/m² 1-6 1-6 2A 500 mg BID: on day 1 for days 8-28 cycles 2-6 for cycle1 days 1-28 for subsequent cycles

Compound 1 besylate, rituximab and bendamustine will be administeredaccording to the cohorts for Arm B of the study, listed in Table 5:

TABLE 5 Study Dosing Schema for Arm B Escalating Dose Portion of StudyCompound 1 COHORT besylate Rituximab Bendamustine 1B 375 mg BID: 6 doses12 doses days 8-28 administered administered for cycle 1 as follows: asfollows: days 1-28 375 mg/m² 70 mg/m² for subsequent on day 1 on days1-2 cycles for cycle 1 for cycles 1-6 2B 500 mg BID: 500 mg/m² days 8-28on day 1 for cycle 1 for cycles 2-6 days 1-28 for subsequent cycles

Within each cohort, subjects will be treated PO (oral) BID (daily) withCompound 1 besylate according to Tables 4 or 5 during two initial 28-daytreatment cycles and will be assessed for safety, tolerability and DLT,as well as pharmacokinetic (“PK”), pharmacodynamic (“PD”), and diseaseresponse.

Rituximab will be administered as a single intravenous (IV) infusion.The initial infusion during cycle 1 will be administered at 375 mg/m²;subsequent infusions during cycles 2 through 6 will be administered at500 mg/m². Administration of rituximab will begin on day 1 of cycle 1and on day 1 of each cycle thereafter. Following the cycle 6 infusion,rituximab will be discontinued. Subjects may continue on treatment withCompound 1 besylate if appropriate. The first infusion of rituximab willbe at a rate of 50 mg/hr. In the absence of infusion toxicity, theinfusion rate will be increased by 50 mg/hr increments every 30 minutes,to a maximum of 400 mg/hr. Each subsequent infusion will be initiated at100 mg/hr. In the absence of infusion toxicity, the infusion rate willbe increased by 100 mg/hr increments at 30 minute intervals to a maximumof 400 mg/hr.

Fludarabine will be administered as a 25 mg/mL (Sterile Water USP)intravenous infusion of 25 mg/m² over 20-30 minutes on days 1-3 forcycles 1-6. Cyclophosphamide will be administered as a 100 mg/5 mLintravenous infusion of 250 mg/m² over 10-30 minutes on days 1-3 forcycles 1-6.

Bendamustine will be diluted to 5 mg/mL concentration with Sterile Waterfor Injection. Immediately prior to use, the bendamustine solution willbe transferred to a 500 mL infusion bag of 0.9% Sodium ChlorideInjection USP. The bendamustine infusion solution will then beadministered as an intravenous infusion of 70 mg/m² over 30-60 minuteson days 1 and 2 for cycles 1-6.

Example 4

Capsules comprisingN-(3-(5-fluoro-2-(4-(2-methoxyethoxy)phenylamino)pyrimidin-4-ylamino)phenyl)acrylamidebesylate (Compound 1 besylate) for use in this study correspond to Table1 in Example 1.

Prior to administration, each dose of ofatumumab is prepared in 1000 mL0.9% Sodium Chloride Injection, USP solutions. The dilution ofofatumumab is as follows:

-   -   300 mg dose: 5 mL from each of 3 single use 100 mg vials is        added to 985 mL of 0.9% Sodium Chloride Injection, USP.    -   2000 mg dose: 50 mL from each of 2 single use 1000 mg vials is        added to 900 mL of 0.9% Sodium Chloride Injection, USP.

Study Design

Subjects with relapsed or refractory CLL or SLL who failed at least oneprior treatment regimen will be enrolled in a “3+3” dose escalation andexpansion study to determine the Not Tolerated Dose (NTD), the OptimalBiologic Effect dose (OBE) and the Maximum Tolerated Dose (MTD) ofCompound 1 and ofatumumab. Approximately 30-42 patients are expected tobe enrolled in the study.

Study treatment will be administered in 28-day cycles at specified doselevels as scheduled until disease progression, unacceptable toxicity, ordiscontinuation for any other reason. Subjects will continue on thestarting dose until the preliminary recommended Phase 2 dose (RP2D) isdetermined, at which point they can be switched to the preliminary RP2D.

Compound 1 besylate and ofatumumab will be administered according to thecohorts for the study, listed in Table 6:

TABLE 6 Study Dosing Schema for Escalating Dose Portion of StudyCompound 1 COHORT besylate Ofatumumab 1 375 mg BID 12 doses administeredas follows: for days 1-28 300 mg initial dose, followed 1 2 500 mg BIDweek later by for days 1-28 2000 mg weekly for 7 doses, followed 4 weekslater by 2000 mg every 4 weeks for 4 doses

Within each cohort, subjects will be treated PO (oral) BID (daily) withCompound 1 besylate according to Table 6 during two initial 28-daytreatment cycles and will be assessed for safety, tolerability and DLT,as well as pharmacokinetic (“PK”), pharmacodynamic (“PD”), and diseaseresponse.

Ofatumumab will be administered as a single intravenous (IV) infusion.The initial infusion during the first dose will be at a rate of 3.6mg/hour (12 mL/hour). The infustion rate of dose 2 will be at a rate of24 mg/hour (12 mL/hour). Subsequent infusion rates will be at 50 mg/hour(25 mL/hour). In the absence of infusional toxicity, the rate ofinfustion may be increased every 30 minutes as described in Table 7:

TABLE 7 Infusion Rates for Ofatumumab Interval After Start of InfusionDose 1^(a) Dose 2^(b) Doses 3-12^(b) (min) (mL/hour) (mL/hour) (mL/hour)0-30 12 12 25 31-60  25 25 50 61-90  50 50 100 91-120 100 100 200 >120200 200 400 ^(a)Dose 1 = 300 mg (0.3 mg/mL) ^(b)Doses 2 and 3-12 = 2000mg (2 mg/mL)

1. A method of treating, stabilizing or lessening the severity orprogression of one or more diseases and conditions associated with BTKcomprising administering to a patient in need thereof an irreversibleBTK inhibitor and an anti-CD20 antibody, wherein the irreversible BTKinhibitor has not more than about 50% inhibition of a kinase selectedfrom c-Kit, PDGFRa, RIPK2, HCK, EPHA6, LYN, CSK, LCK, ZAK/MLTK, LYN B,FRK/PTK5, FYN, BRAF, RIPK3, ARAF and SRMS, or combinations thereof. 2.The method according to claim 1, wherein the irreversible BTK inhibitorhas not more than about 30% inhibition of a kinase selected from c-Kit,RIPK2, HCK, EPHA6, LYN, CSK, ZAK/MLTK, LYN B, FRK/PTK5, FYN, BRAF,RIPK3, ARAF and SRMS, or combinations thereof.
 3. The method accordingto claim 1, wherein the irreversible BTK inhibitor has not more thanabout 10% inhibition of a kinase selected from EPHA6, LYN B, FRK/PTK5,BRAF, RIPK3, ARAF and SRMS, or combinations thereof.
 4. The methodaccording to claim 1, wherein the irreversible BTK inhibitor has apercent inhibition of LYN that is not more than about 20-30%.
 5. Themethod according to claim 1, wherein the anti-CD20 antibody is selectedfrom rituximab and ofatumumab. 6-22. (canceled)
 23. A system fortreating, stabilizing or lessening the severity of one or more diseasesor conditions associated with BTK, the system comprising Compound 1, ora pharmaceutically acceptable salt thereof, an anti-CD20 antibody and atleast one additional therapeutic agent selected from fludarabine,cyclophosphamide and bendamustine.
 24. The system according to claim 23,wherein the anti-CD20 antibody is selected from rituximab andofatumumab.